[Clinical application of non-invasive prenatal testing for twin pregnancies].

OBJECTIVE: To evaluate the feasibility of non-invasive prenatal testing (NIPT) for the screening of fetal chromosome aneuploidies in twin pregnancies. METHODS: A total of 2 745 women with twin-pregnancies were subjected for NIPT screening. Chromosomal karyotyping and chromosomal microarray analysis (CMA) were carried out on amniotic fluid samples from those with a high risk for fetal chromosome aneuploidies, and the diagnosis and pregnancy outcome were followed up. The sensitivity, specificity, positive predictive value and false positive rate of the NIPT were calculated. RESULTS: Compared with other chromosomal abnormalities, NIPT had a higher efficacy for trisomy 21 and sex chromosomal aneuploidy (SCA) in twin pregnancies (with sensitivity being 100%, 100%, and specificity being 99.93%, 99.9%, respectively). It is difficult to evaluate the efficacy for trisomies 18 and 13 due to the limited data. For chromosome microdeletions and microduplications spanning 15 ~ 21 Mb, NIPT also had a certain detection rate. Compared with women with natural conception, NIPT had a higher detection rate for those with twin pregnancies by assisted reproduction (P < 0.05). CONCLUSION: It is feasible to use NIPT for the detection of chromosome aneuploidies in women with twin pregnancies.

α-Helix-Mediated Protein Adhesion.

Proteins have been adopted by natural living organisms to create robust bioadhesive materials, such as biofilms and amyloid plaques formed in microbes and barnacles. In these cases, ß-sheet stacking is recognized as a key feature that is closely related to the interfacial adhesion of proteins. Herein, we challenge this well-known recognition by proposing an α-helix-mediated interfacial adhesion model for proteins. By using bovine serum albumin (BSA) as a model protein, it was discovered that the reduction of disulfide bonds in BSA results in random coils from unfolded BSA dragging α-helices to gather at the solid/liquid interface (SLI). The hydrophobic residues in the α-helix then expose and break through the hydration layer of the SLI, followed by the random deposition of hydrophilic and hydrophobic residues to achieve interfacial adhesion. As a result, the first assembled layer is enriched in the α-helix secondary structure, which is then strengthened by intermolecular disulfide bonds and further initiates stepwise layering protein assembly. In this process, ß-sheet stacking is transformed from the α-helix in a gradually evolving manner. This finding thus indicates a valuable clue that ß-sheet-featuring amyloid may form after the interfacial adhesion of proteins. Furthermore, the finding of the α-helix-mediated interfacial adhesion model of proteins affords a unique strategy to prepare protein nanofilms with a well-defined layer number, presenting robust and modulable adhesion on various substrates and exhibiting good resistance to acid, alkali, organic solvent, ultrasonic, and adhesive tape peeling.

Transesophageal electrophysiology study in the diagnosis of dual atrioventricular nodal nonreentrant tachycardia.

"Double fire" is generally characterized by 1:2 atrioventricular conduction of sinus beats traveling down fast and slow pathways that result in double ventricular response. When this phenomenon repeats rapidly, dual atrioventricular nodal nonreentrant tachycardia (DAVNNT) occurs. We report a case of an irregular tachycardia with a comprehensive record that includes an electrocardiogram, a transesophageal electrophysiology study, and an intracardiac electrophysiology study. This is the first report of transesophageal electrophysiology study in the diagnosis of DAVNNT. A diagnosis of DAVNNT was deduced, and the patient was successfully treated with radiofrequency ablation of the slow pathway.

Efficient and genotype independent maize transformation using pollen transfected by DNA-coated magnetic nanoparticles.

Current gene delivery methods for maize are limited to specific genotypes and depend on time-consuming and labor-intensive tissue culture techniques. Here, we report a new method to transfect maize that is culture-free and genotype independent. To enhance efficiency of DNA entry and maintain high pollen viability of 32%-55%, transfection was performed at cool temperature using pollen pretreated to open the germination aperture (40%-55%). Magnetic nanoparticles (MNPs) coated with DNA encoding either red fluorescent protein (RFP), ß-glucuronidase gene (GUS), enhanced green fluorescent protein (EGFP) or bialaphos resistance (bar) was delivered into pollen grains, and female florets of maize inbred lines were pollinated. Red fluorescence was detected in 22% transfected pollen grains, and GUS stained 55% embryos at 18 d after pollination. Green fluorescence was detected in both silk filaments and immature kernels. The T1 generation of six inbred lines showed considerable EGFP or GUS transcripts (29%-74%) quantitated by polymerase chain reaction, and 5%-16% of the T1 seedlings showed immunologically active EGFP or GUS protein. Moreover, 1.41% of the bar transfected T1 plants were glufosinate resistant, and heritable bar gene was integrated into the maize genome effectively as verified by DNA hybridization. These results demonstrate that exogenous DNA could be delivered efficiently into elite maize inbred lines recalcitrant to tissue culture-mediated transformation and expressed normally through our genotype-independent pollen transfection system.

Transcriptional mutagenesis mediated by 8-oxoG induces translational errors in mammalian cells.

Reactive oxygen species formed within the mammalian cell can produce 8-oxo-7,8-dihydroguanine (8-oxoG) in mRNA, which can cause base mispairing during gene expression. Here we found that administration of 8-oxoGTP in MTH1-knockdown cells results in increased 8-oxoG content in mRNA. Under this condition, an amber mutation of the reporter luciferase is suppressed. Using second-generation sequencing techniques, we found that U-to-G changes at preassigned sites of the luciferase transcript increased when 8-oxoGTP was supplied. In addition, an increased level of 8-oxoG content in RNA induced the accumulation of aggregable amyloid ß peptides in cells expressing amyloid precursor protein. Our findings indicate that 8-oxoG accumulation in mRNA can alter protein synthesis in mammalian cells. Further work is required to assess the significance of these findings under normal physiological conditions.

Risk factors for treatment failure of heated humidified high-flow nasal cannula as initial respiratory support for preterm infants. / æ—©äº§å„¿åŠ æ¸©æ¹¿åŒ–é«˜æµé‡ç»é¼»å¯¼ç®¡é€šæ°”åˆå§‹å‘¼å¸æ”¯æŒå¤±è´¥çš„å±é™©å› ç´ åˆ†æž.

OBJECTIVES: To study the risk factors for treatment failure of heated humidified high-flow nasal cannula (HHHFNC) as initial respiratory support for preterm infants. METHODS: A retrospective analysis was performed on the medical data of the preterm infants who were admitted from January 2018 to April 2021 and received HHHFNC for initial respiratory support after birth. According to whether it was necessary to upgrade to noninvasive continuous positive airway pressure or invasive mechanical ventilation within 72 hours after treatment, they were divided into a failure group and a success group. Univariate and multivariate logistic regression analyses were used to determine the risk factors for failure of HHHFNC as initial respiratory support. RESULTS: A total of 166 preterm infants were included, among whom 48 (28.9%) experienced the treatment failure of HHHNFC as initial respiratory support. The univariate analysis showed that compared with the success group with 118 infants, the failure group had significantly lower gestational age and birth weight and a significantly higher proportion of infants with fraction of inspired oxygen >35%, flow rate >6 L/minute, patent ductus arteriosus (PDA), respiratory distress syndrome (RDS), or use of pulmonary surfactant (P<0.05). The multivariate logistic regression analysis showed that gestational age <32 weeks, PDA (>1.5 mm and left atrium/aorta diameter ratio >1.4), fraction of inspired oxygen >35%, flow rate >6 L/minute, and presence of RDS were risk factors for the treatment failure of HHHNFC as initial respiratory support (P<0.05). CONCLUSIONS: The preterm infants with a gestational age of <32 weeks or the presence of RDS tend to have a high risk of failure of HHHNFC as initial respiratory support. The risk of failure of HHHFNC as initial respiratory support increases in infants with oxygen concentration >35% and/or flow rate >6 L/minute, or the presence of PDA, suggesting an upgrade of respiratory support should be considered. Citation.

Influences of age, gender, and circadian rhythm on deceleration capacity in subjects without evident heart diseases.

BACKGROUND: Deceleration capacity (DC) is a newly found predictor of mortality after myocardial infarction. Age-, gender-, and circadian rhythm-related differences in DC may limit its predictive value, which should be considered in clinical settings. METHODS: DC, average heart rate, and HRV parameters, including 24 hours, awaking state (15:00-20:00) and sleeping mode (00:00-05:00) strips from 24 hours Holter recordings in 636 subjects without heart diseases were examined. Heart rate variability was analyzed in time domains (standard deviation of all normal-to-normal intervals [SDNN], normal-to-normal RR intervals in all 5-minute segments [SDANN], and root mean square successive difference [RMSSD]). RESULTS: The DC, SDNN, SDANN, RMSSD, and heart rate decreased with age. Deceleration capacity was significantly lower in patients greater than 50 years of age. The largest decrease of SDNN, SDANN, and RMSSD occurred in patients 30-39 years of age. The values of SDNN, SDANN, and DC of women were lower than that of men in the young and middle-aged groups, but age-related decrease of DC in men was greater than that in women. Heart rate of women was significantly higher than that of men in younger subjects, especially in a sleeping mode. There were higher values of DC and RMSSD during sleeping than that during a waking state. CONCLUSIONS: The age, gender, and circadian rhythm may be useful when evaluating cardiac autonomic function and need to be considered when evaluating DC and HRV in clinical and scientific researches.

Neural regeneration: role of traditional Chinese medicine in neurological diseases treatment.

The effects of a single compound and a mixture of traditional Chinese medicine (TCM) on promoting proliferation, differentiation, and migration of neural stem cells and regulating their microenvironment have been observed by Chinese scholars in recent years. These results showed good prospects in improving neural regeneration and repair of neurological disorders such as ischemic brain injury, Alzheimer's disease, Parkinson's disease, and depression. According to the TCM theory, the relationship between life of an individual and the disease was regarded as an entirety, and the theory emphasized the treatment based on syndrome differentiation since ancient times. In this paper, we attempted to introduce these medicines, which belong to natural products and have already been proved to possess clear therapeutic action on human bodies in the clinical setting. We summarized their effects promoting brain neurogenesis and repairing brain injuries in animal models and some mechanisms at the cellular and molecular levels.

CD151 promotes proliferation and migration of PC3 cells via the formation of CD151-integrin α3/α6 complex.

Over-expression of CD151 was found to be associated with metastasis and poor prognosis of prostatic carcinoma. This study was designed to examine the mechanism by which CD151 promotes the proliferation and migration of prostatic cancer cells. The pAAV-CD151, pAAV-GFP and pAAV-CD151-AAA mutant plasmids were constructed and used to transiently transfect PC3 cells (a prostatic carcinoma 3 cell line) by the mediation of Fugene HD. Then, the cells were assigned to control group, pAAV-GFP group, pAAV-CD151 group, and pAAV-CD151-AAA group respectively. Cell proliferation was evaluated by using the 3-[4,5-dimet-hylthiazol-2-yl]-2,5, diphenyltetrazolium bromide (MTT) method. Cell migration assay was performed by using Boyden chambers. The formation of CD151-integrin α3/α6 complex was determined by the method of co-immunoprecipitation. The protein expression levels of CD151 and extracellular signal-regulated kinase (ERK) were measured by Western blotting. The results showed that transfection of pAAV-CD151 or pAAV-CD151-AAA mutant increased the expression of CD151 protein in PC3 cells. Co-immunoprecipitation showed that more CD151-integrin α3/α6 complex was formed in the pAAV-CD151 group than in the control group, the pAAV-GFP group and the pAAV-CD151-AAA mutant group. Furthermore, the proliferative and migrating capacity of PC3 cells was substantially increased in the pAAV-CD151 group but inhibited in the pAAV-CD151-AAA mutant group. CD151 transfection increased the expression of phospho-ERK. Taken together, it was concluded that CD151 promotes the proliferation and migration of PC3 cells through the formation of CD151-integrin complex and the activation of phosphorylated ERK.

Coupling isotopic analysis and Ecopath model to detect the ecosystem features based on functional groups of the southwestern Yellow Sea, China.

The paper evaluates the species richness, material transfer, energy flow, and system function of the southwestern Yellow Sea Ecosystem (SYSE) indicating intensive human intervention affecting this large marine ecosystem. Twenty functional groups were chosen to represent the basic components of the SYSE for Ecopath modeling based on offshore surveys, annual bird observations, and the China Fisheries Statistical Yearbooks. Forty-nine species based on 15 functional groups of Ecopath model were assessed by stable isotopic analysis (SIA) to verify ecosystem features, energy flow, and trophic structure of the SYSE derived from Ecopath model. Results showed there was a clear correlation of the estimated trophic structure calculated from SIA and the Ecopath model with R2=0.7184. The SYSE Ecopath model was still at an immature and unstable stage according to outputs of the modeling parameters. This paper provides a verification method of detecting the ecosystem features and maturity, stability, and resilience of marine ecosystems by comparing outputs from Ecopath models with SIA.

Determining nitrate sources in storm runoff in complex urban environments based on nitrogen and oxygen isotopes.

Urban storm runoff, as the primary transport medium for nutrients entering urban rivers, contributes to urban water contamination. Accurate source identification is critical for controlling water pollution. Although some studies have used nitrate isotopic composition (δ15N-NO3- and δ18O-NO3-) to identify nitrate (NO3--N) in urban storm runoff, the relatively low frequency of collecting samples in surface runoff within a single functional area hinders the understanding of spatial variations and dynamic process of NO3--N sources over the runoff process. This study investigated the nitrogen (N) concentrations and analyzed dynamic changes of NO3--N sources in surface runoff in different urban functional areas, drainage pipeline runoff, and channels during the complete runoff process in Wuxi, east China. The results showed that N concentrations in pipeline runoff and channels were higher than those in surface runoff, indicating that high concentration of N pollutants were accumulated in drainage pipelines. Information of δ15N-NO3- and δ18O-NO3- suggested that the main NO3--N source varied between runoff stages. NO3--N contribution from atmospheric deposition decreased in the order: surface runoff (57%) > residential pipeline runoff (25%) > channels (14%), while the opposite trend was observed for the contributions from sewage, increasing from 10%, 26% to 39%. In urban storm runoff, more sewage, fertilizers, and soil N were carried into the surface runoff after 30% of cumulative runoff ratio and carried into pipeline runoff in the initial 25% of cumulative runoff ratio in the residential area. As the first attempt to identify nitrate sources over the cumulative runoff in different urban functional areas, this work expands our understanding of the primary nitrate source in urban storm runoff. The findings provide important insights for developing strategies to mitigate non-point source water pollution.

Coronavirus disease 2019 death prediction by electrocardiographic abnormalities and elevated D-dimer levels.

Background: Electrocardiography (ECG) plays a very important role in various cardiovascular diseases and elevated D-dimer in serum associated with thrombosis. In patients with coronavirus disease 2019 (COVID-19), immense pieces of evidence showed that ECG abnormalities or elevated D-dimer in serum occurred frequently. However, it remains unclear whether ECG abnormalities combined with elevated D-dimer could be a new risk predictor in patients with COVID-19. Methods and results: This retrospective cohort study enrolled 416 patients with COVID-19 at Wuhan Tongji Hospital from 1 February to 20 March 2020. ECG manifestations, D-dimer levels, and in-hospital deaths were recorded for all patients. Logistic regression analysis was performed to examine the association between ECG manifestations and in-hospital mortality in patients with elevated D-dimer levels. In patients hospitalized for COVID-19, ST-T abnormalities (34.3%) were the most frequent ECG manifestations, whereas sinus tachycardia (ST) (13.3%) and atrial arrhythmias with rapid rhythms (8.5%) were the two most common cardiac arrhythmias. Compared to severely ill patients with COVID-19, ST-T abnormalities, ST and atrial arrhythmias (p<0.001) with rapid rhythms, D-dimer levels, and in-hospital deaths were significantly more frequent in critically ill patients with COVID-19. Moreover, elevated D-dimer levels were observed in all the patients who died. In the subgroup of 303 patients with elevated serum D-dimer levels, the patient's age, the incidence of ST-T abnormalities, ST, atrial fibrillation (AF), and atrial premature beat were significantly higher than those in the non-elevated D-dimer subgroup. Multivariate logistic regression analysis further revealed that ST and AF were risk factors for in-hospital mortality in COVID-19 patients with elevated D-dimer levels. Conclusions: ECG abnormalities and elevated D-dimer levels were associated with a higher risk of critical illness and death in patients hospitalized for COVID-19. ECG abnormalities, including ST and AF, combined with elevated D-dimer levels, can be used to predict death in COVID-19.

Quantitative identification of non-point sources of nitrate in urban channels based on dense in-situ samplings and nitrate isotope composition.

Quantitative identification of non-point sources of nitrate in urban channels plays a critical role in effective nutrient management in urban regions. This is an emerging issue due to fast urbanization and the resultant complicated hydrological and hydraulic conditions in urban areas. In this study, we examine spatial-temporal characteristics of nitrogen concentration in urban channels based on dense in-situ samplings during a one-year period over a small urban catchment in China. We quantitatively identify nitrate sources into urban channels based on dual-isotope analyses and Bayesian isotope mixing model. Results show that nitrogen concentration peaks in winter as well as in urban channels and land surfaces in the urban core region. Sewage (47%) is the dominate contributor to NO3--N in urban channels, followed by NH4+ in fertilizer (30%) as the second contributor. Sewage (NH4+ in fertilizer) contributes more NO3--N to channels in winter (summer) with the proportion of 65% (44%), and more NO3--N to urban core (suburban) channels with the proportion of 59% (42%). The rainfall and distribution of rainwater drains explain the monthly and spatial variations of contribution of NO3--N sources well, respectively. In addition, less NO3--N in the urban channels derives from nitrification, which is consistent with the results of high properties of NH4+-N/TN in this region. Our results highlight the key roles of land use types and rainfall in NO3--N source apportionment, and provide support for the nitrogen management practices in urbanized regions.

Protective effects of octacosanol on 6-hydroxydopamine-induced Parkinsonism in rats via regulation of ProNGF and NGF signaling.

AIM: To investigate the protective effects of octacosanol in 6-hydroxydopamine-induced Parkinsonian rats and find whether octacosanol has effects on pro nerve growth factor (pro-NGF), NGF and the downstream effector proteins. METHODS: Behavioral tests, enzymatic assay, tyrosine hydroxylase immunohistochemistry, TUNEL and Western blot were used to investigate the effects of octacosanol in this rat model of PD. RESULTS: Oral administration of octacosanol (35-70 mg/kg, po for 14 d) significantly improved the behavioral impairments in rats induced by 6-OHDA and dose-dependently preserved the free radical scavenging capability of the striatum. Octacosanol treatment also effectively ameliorated morphological appearances of TH-positive neuronal cells in nigrostriatal systems and decreased the apoptotic cells induced by 6-OHDA in striatum. In addition, octacosanol strikingly blocked the 6-OHDA-induced increased expression of proNGF-p75NTR-sortilin death signaling complex and its downstream effector proteins. Meantime, octacosanol prevented the decreased levels of NGF, its receptors TrkA and p-Akt which together mediated the cell survival pathway. CONCLUSION: The findings implicated that the anti-parkinsonism effects afforded by octacosanol might be mediated by its neuro-microenvironment improving potency through retrieving the ratios of proNGF:NGF and the respective receptors p75NTR:TrkA in vivo. Due to its excellent tolerability and non-toxicity, octacosanol may be a promising agent for PD treatment.

[Relationship of oxidative DNA damage and expression of mitochondrial apoptotic proteins in rat striatum induced by 6-hydroxydopamine].

OBJECTIVE: To study the relationship between oxidative DNA damage and mitochondrial apoptosis relative proteins in rat striatum induced by 6-hydroxydopamine (6-OHDA) during the pathogenesis of Parkinson's disease (PD). METHODS: 6-OHDA was stereotactically injected into the bilateral right striatum of rats to produce PD models. Assays for 8-oxo-dG immunohistochemistry and Western blot for MTH1, Cytochrome c, Cl-caspase 9 and Cl-caspase 3 in right striatum was separately conducted. RESULTS: In 10 successful PD rats, compared with either sham or normal group, there were obvious more 8-oxo-dG positive cells in lesioned striatum while there was a lower expression of MTH1. Furthermore, the expressions of such intrinsic apoptotic pathway factors as cytoplasmic Cytochrome c, Cl-caspase 9 and Cl-caspase 3 were highly up-regulated in lesioned striatum. CONCLUSION: Oxidative DNA damage plays a key role in the pathogenesis of PD. Furthermore Cytochrome c, caspase 9 and caspase 3 are involved in the regulation of apoptosis under oxidative DNA damage induced by 6-OHDA.

Zeolite-Enhanced Sustainable Pd-Catalyzed C-C Cross-Coupling Reaction: Controlled Release and Capture of Palladium.

Supported palladium catalysts have attracted significant attention for use in cross-coupling reactions due to their recyclability. However, the inevitable progressive loss of Pd that occurs in the catalytic process deactivates the catalysts, which hinders their sustainable application. Herein, we report a zeolite-enhanced sustainable Pd catalyst for C-C cross-coupling reactions. Zeolite does a good job of acting as a sink for Pd2+ ions. This catalyst exhibits an excellent homogeneous catalytic performance by releasing Pd species from zeolite. In addition, the Pd2+ ions were successfully recaptured in a controlled catalytic system by combining the uniform microporous structure and good adsorption features of zeolite. The release/capture mechanism of the Pd species guaranteed the high loading and high dispersion of Pd on the recycled catalyst. The 0.84%Pd@USY catalysts were reused at least 10 times in water without an appreciable reduction in activity. This study presents a new perspective toward the design of a highly efficient and sustainable supported metal catalyst.

The complete genome sequence of the algicidal bacterium Bacillus subtilis strain JA and the use of quorum sensing to evaluate its antialgal ability.

We describe the isolation of Bacillus subtilis strain JA and demonstrate that this bacterium exhibited strong algicidal effects on the algae Alexandrium minutum with an inhibition rate exceeding 80 % within 48 h. B. subtilis JA significantly reduced the photosynthetic efficiency of A. minutum and caused extensive morphological damage to the algae. Genomic analysis of B. subtilis JA demonstrated that a putative AI-2 type quorum sensing (QS) gene (LuxS) is present in its genome cluster, which is regulate pheromone biosynthesis. Interestingly, the exogenous addition of a QS-oligopeptide (ComX-pheromone) improved the algicidal efficiency of B. subtilis JA, thus indicating that the algicidal activity of this bacterium is potentially regulated by QS. Collectively, our data describe a potential antialgal bacterium and speculated that its behavior can be modulated by QS signal. B. subtilis JA may therefore represent a valuable tool for the development of novel chemical-ecological methods with which to control harmful algae.

Temporal Variability of Virioplankton during a Gymnodinium catenatum Algal Bloom.

Viruses are key biogeochemical engines in the regulation of the dynamics of phytoplankton. However, there has been little research on viral communities in relation to algal blooms. Using the virMine tool, we analyzed viral information from metagenomic data of field dinoflagellate (Gymnodinium catenatum) blooms at different stages. Species identification indicated that phages were the main species. Unifrac analysis showed clear temporal patterns in virioplankton dynamics. The viral community was dominated by Siphoviridae, Podoviridae, and Myoviridae throughout the whole bloom cycle. However, some changes were observed at different phases of the bloom; the relatively abundant Siphoviridae and Myoviridae dominated at pre-bloom and peak bloom stages, while at the post-bloom stage, the members of Phycodnaviridae and Microviridae were more abundant. Temperature and nutrients were the main contributors to the dynamic structure of the viral community. Some obvious correlations were found between dominant viral species and host biomass. Functional analysis indicated some functional genes had dramatic response in algal-associated viral assemblages, especially the CAZyme encoding genes. This work expands the existing knowledge of algal-associated viruses by characterizing viral composition and function across a complete algal bloom cycle. Our data provide supporting evidence that viruses participate in dinoflagellate bloom dynamics under natural conditions.

Complete genome sequence of Acinetobacter baumanni J1, a quorum sensing-producing algicidal bacterium, isolated from Eastern Pacific Ocean.

The Acinetobacter baumanni J1 isolated from surface water of the Eastern Pacific Ocean, demonstrated significant algicidal activity on the algae Alexandrium tamarense. Interestingly, this strain showed the ability to produce an acyl-homoserine lactone (AHL) quorum sensing molecule. To better understand its AHL producing mechanism and its ecological functions, the genome of A. baumanni strain J1 was completely sequenced. The genome contained a circular chromosome of 3,948,465 bp with an average GC content of 39.9 mol%. A total of 3707 protein coding genes, 41 tRNA genes and 16 rRNA genes were obtained. In silico genome annotation identified a LuxI putative gene located on contig 4. Subsequent thin-layer chromatography analysis indicated that C8-AHL could be produced by A. baumanni J1, which confirmed the authenticity of the LuxI gene. Taken together, this work describes an algicidal bacterium that is capable of producing an AHL molecule, which may represent a valuable tool for developing microbial methods to control harmful algae.