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1.
Gen Hosp Psychiatry ; 83: 86-92, 2023 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-37148598

RESUMO

OBJECTIVE: Multiple studies have indicated that electroconvulsive therapy (ECT) could increase brain-derived neurotrophic factor (BDNF) concentrations in patients with different mental disorders. The aim of this synthesis was to evaluate post-ECT BDNF concentrations in patients with various mental disorders. METHODS: The Embase, PubMed and Web of Science databases were systematically searched for studies in English comparing BDNF concentrations before and after ECT through 11/2022. We extracted the pertinent information from the included studies and evaluated their quality. The standardized mean difference (SMD) with a 95% confidence interval (CI) was calculated to quantify BDNF concentration differences. RESULTS: In total, 35 studies assessed BDNF concentrations in 868 and 859 patients pre and post-ECT treatment, respectively. Post-ECT-treatment BDNF concentrations were significantly higher than the pretreatment concentrations (Hedges'g = -0.50, 95% CI (-0.70, -0.30), heterogeneity I2 = 74%, p < 0.001). The analysis that combined both ECT responders and non-responders demonstrated a marked increase in total BDNF levels subsequent to ECT treatment (Hedges'g = -0.27, 95% CI (-0.42, -0.11), heterogeneity I2 = 40%, p = 0.0007). CONCLUSION: Irrespective of the effectiveness of ECT, Our study shows that peripheral BDNF concentrations increase significantly after the entire course of ECT, which may enhance our comprehension of the interplay between ECT treatment and BDNF levels. However, BDNF concentrations were not associated with the effectiveness of ECT, and abnormal concentrations of BDNF may be linked to the pathophysiological process of mental illness, necessitating more future research.

2.
Brain ; 2023 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-37137515

RESUMO

Human speech and language are among the most complex motor and cognitive abilities. The discovery of a mutation in the transcription factor FOXP2 in KE family members with speech disturbances has been a landmark example of the genetic control of vocal communication in humans. Cellular mechanisms underlying this control have remained unclear. By leveraging FOXP2 mutation/deletion mouse models, we found that the KE family FOXP2R553H mutation directly disables intracellular dynein-dynactin 'protein motors' in the striatum by induction of a disruptive high level of dynactin1 that impairs TrkB endosome trafficking, microtubule dynamics, dendritic outgrowth and electrophysiological activity in striatal neurons alongside vocalization deficits. Dynactin1 knockdown in mice carrying FOXP2R553H mutations rescued these cellular abnormalities and improved vocalization. We suggest that FOXP2 controls vocal circuit formation by regulating protein motor homeostasis in striatal neurons, and that its disruption could contribute to the pathophysiology of FOXP2 mutation/deletion-associated speech disorders.

4.
ASAIO J ; 69(5): 490-495, 2023 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-37126229

RESUMO

This study evaluated the suitability, feasibility, safety, and outcomes of transport of the ECMO-dependent patient (EDP) by EDP transport team (EDPTT) in China. Eighty-two EDPs (forty-one cases on VV ECMO and forty-one cases on VA ECMO) received transport between June 2018 and June 2021 and were retrospectively analyzed. ECMO circulation was performed by the outlying hospital, mainly using percutaneous ECMO cannulation. The EDPTT consists of three intensive therapists, one of whom serves as a team leader, and one intensive care unit nurse. Of these, 81 (98.8%) patients were transferred by ambulance, no deaths occurred during transport, the EDP-related complications were 19% (n = 16); bleeding at the cannula site (n = 7, 8.5%) was the most prominent; equipment-related problems accounted for 14.6% of the problems requiring urgent intervention, with hand cranking being the most common (9.7%). The survival rate during transport was 100%, with 36 (43.9%) patients surviving to discharge. The ECMO weaning rate was 61% for VV ECMO and 63.7% for VA ECMO. The results demonstrated the suitability, feasibility, and safety of transporting EDP in a team led by an intensivist, with few complications and no deaths during transport. This may be the recommended staffing model for EDP transport in developing countries.


Assuntos
Oxigenação por Membrana Extracorpórea , Humanos , Oxigenação por Membrana Extracorpórea/métodos , Transporte de Pacientes/métodos , Estudos Retrospectivos , Unidades de Terapia Intensiva , Transferência de Pacientes/métodos
5.
Materials (Basel) ; 16(9)2023 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-37176205

RESUMO

The adsorption, dissociation and penetration processes of N2 on the surface of ZrMnFe(110) were investigated using the first-principles calculation method in this paper. The results indicate that the vacancy Hollow 1 composed of 4Zr1Fe on the surface of ZrMnFe(110) is the best adsorption site for the N2 molecule and N atom, and the adsorption energies are 10.215 eV and 6.057 eV, respectively. Electron structure analysis indicates that the N2 molecule and N atoms adsorbed mainly interact with Zr atoms on the surface. The transition state calculation shows that the maximum energy barriers to be overcome for the N2 molecule and N atom on the ZrMnFe(110) surface were 1.129 eV and 0.766 eV, respectively. This study provides fundamental insight into the nitriding mechanism of nitrogen molecules in ZrMnFe.

6.
Sensors (Basel) ; 23(9)2023 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-37177763

RESUMO

Soft biological tissues perform various functions. Sensory nerves bring sensations of light, voice, touch, pain, or temperature variation to the central nervous system. Animal senses have inspired tremendous sensors for biomedical applications. Following the same principle as photosensitive nerves, we design flexible ionic hydrogels to achieve a biologic photosensor. The photosensor allows responding to near-infrared light, which is converted into a sensory electric signal that can communicate with nerve cells. Furthermore, with adjustable thermal and/or electrical signal outputs, it provides abundant tools for biological regulation. The tunable photosensitive performances, high flexibility, and low cost endow the photosensor with widespread applications ranging from neural prosthetics to human-machine interfacing systems.


Assuntos
Biônica , Percepção do Tato , Animais , Humanos , Hidrogéis , Tato , Neurônios
7.
Artigo em Inglês | MEDLINE | ID: mdl-37159591

RESUMO

Liang-Ge (LG) decoction could ameliorate coagulation dysfunction in septic model rats. However, the mechanism of LG in treating sepsis still needs to be clarified. Our current study established a septic rat model to evaluate the effect of LG on coagulation dysfunction in septic rats first. Second, we investigated the effect of LG on NET formation in septic rats. Finally, NETs and PAD4 inhibitors were further used to clarify if LG could improve the mechanism of sepsis coagulation dysfunction by inhibiting NET formation. Our findings indicated that treatment with LG improved the survival rate, reduced inflammatory factor levels, enhanced hepatic and renal function, and reduced pathological changes in rats with sepsis. LG could also alleviate coagulation dysfunction in septic model rats. Besides, LG treatment reduced NETs formation and decreased PAD4 expression in neutrophiles. In addition, LG treatment showed a similar result in comparison to the treatment with either NET inhibitors or PAD4 inhibitors alone. In conclusion, this study confirmed that LG has therapeutic effects on septic rats. Furthermore, the improvement of coagulation dysfunction in septic rats by LG was achieved through inhibiting PAD4-mediated NET formation.

8.
Genome Med ; 15(1): 31, 2023 05 02.
Artigo em Inglês | MEDLINE | ID: mdl-37131267

RESUMO

BACKGROUND: Fumarate hydratase-deficient renal cell carcinoma (FH-RCC) is a rare highly aggressive subtype of kidney cancer for which the distinct genomic, transcriptomic, and evolutionary relationships between metastatic and primary lesions are still unclear. METHODS: In this study, whole-exome, RNA-seq, and DNA methylation sequencing were performed on primary-metastatic paired specimens from 19 FH-RCC cases, including 23 primary and 35 matched metastatic lesions. Phylogenetic and clonal evolutionary analyses were used to investigate the evolutionary characteristics of FH-RCC. Transcriptomic analyses, immunohistochemistry, and multiple immunofluorescence experiments were performed to identify the tumor microenvironmental features of metastatic lesions. RESULTS: Paired primary and metastatic lesions generally showed similar characteristics of tumor mutation burden, tumor neoantigen burden, microsatellite instability score, CNV burden, and genome instability index. Notably, we identified an FH-mutated founding MRCA (the most recent common ancestor) clone that dominated the early evolutionary trajectories in FH-RCC. Although both primary and metastatic lesions manifested high immunogenicity, metastatic lesions exhibited higher enrichment of T effector cells and immune-related chemokines, together with upregulation of PD-L1, TIGIT, and BTLA. In addition, we found that concurrent NF2 mutation may be associated with bone metastasis and upregulation of cell cycle signature in metastatic lesions. Furthermore, although in FH-RCC metastatic lesions in general shared similar CpG island methylator phenotype with primary lesions, we found metastatic lesions displaying hypomethylated chemokine and immune checkpoints related genomic loci. CONCLUSIONS: Overall, our study demonstrated the genomic, epigenomic, and transcriptomic features of metastatic lesions in FH-RCC and revealed their early evolutionary trajectory. These results provided multi-omics evidence portraying the progression of FH-RCC.


Assuntos
Carcinoma de Células Renais , Neoplasias Renais , Humanos , Carcinoma de Células Renais/genética , Fumarato Hidratase/genética , Fumarato Hidratase/metabolismo , Transcriptoma , Filogenia , Neoplasias Renais/genética , Neoplasias Renais/patologia , Genômica
9.
Appl Opt ; 62(7): 1662-1666, 2023 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-37132910

RESUMO

Photonic antennas are critical in applications such as spectroscopy, photovoltaics, optical communications, holography, and sensors. Metal antennas are widely used because of their small size, but they are difficult to be compatible with a CMOS. All-dielectric antennas are easier to integrate with Si waveguides, but are generally larger in size. In this paper, we propose the design of a small-sized, high-efficiency semicircular dielectric grating antenna. The antenna's key size is only 2.37µm×4.74µm, and the emission efficiency reaches over 64% in the wavelength range from 1.16 to 1.61 µm. The antenna provides a new, to the best of our knowledge, approach for three-dimensional optical interconnections between different decks of integrated photonic circuits.

10.
Int Immunopharmacol ; 120: 110290, 2023 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-37216800

RESUMO

BACKGROUND: Alzheimer's disease (AD) is a common neurodegenerative disease and a momentous cause of dementia in the elderly. Sennoside A (SA) is an anthraquinone compound and possesses decisive protective functions in various human diseases. The purpose of this research was to elucidate the protective effect of SA against AD and investigate its mechanism. METHODS: Male APPswe/PS1dE9 (APP/PS1) transgenic mice with a C57BL/6J background were chosen as AD model. Age-matched nontransgenic littermates (C57BL/6 mice) were negative controls. SA's functions in AD in vivo were estimated by cognitive function analysis, Western blot, hematoxylin-eosin staining, TUNEL staining, Nissl staining, detection of Fe2+ levels, glutathione and malondialdehyde contents, and quantitative real-time PCR. Also, SA's functions in AD in LPS-induced BV2 cells were examined using Cell Counting Kit-8 assay, flow cytometry, quantitative real-time PCR, Western blot, enzyme-linked immunosorbent assay, and analysis of reactive oxygen species levels. Meanwhile, SA's mechanisms in AD were assessed by several molecular experiments. RESULTS: Functionally, SA mitigated cognitive function, hippocampal neuronal apoptosis, ferroptosis, oxidative stress, and inflammation in AD mice. Furthermore, SA reduced BV2 cell apoptosis, ferroptosis, oxidative stress, and inflammation induced by LPS. Rescue assay revealed that SA abolished the high expressions of TRAF6 and p-P65 (NF-κB pathway-related proteins) induced by AD, and this impact was reversed after TRAF6 overexpression. Conversely, this impact was further enhanced after TRAF6 knockdown. CONCLUSIONS: SA relieved ferroptosis, inflammation and cognitive impairment in aging mice with AD through decreasing TRAF6.

11.
Transl Oncol ; 34: 101683, 2023 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-37224767

RESUMO

BACKGROUND: ALKBH5 belongs to the ALKB family consists of a Fe (II) and a-ketoglutarate-dependent dioxygenase. ALKBH5 directly catalyzes the oxidative demethylation of m6A-methylated adenosine. ALKBH5 involves in tumorigenesis and tumor progression, and is often dysregulated in a wide range of cancers, including colorectal cancer. Emerging evidence indicates that the expression of ALKBH5 is associated with the abundance of infiltrating immune cells in the microenvironment. However, how ALKBH5 affects immune cell infiltration in the microenvironment in colorectal cancer (CRC) has not been reported. The aim of this study was to identify how the expression of ALKBH5 affects the biological behaviors of CRC cell lines and regulates the effects on infiltrating CD8+ T cells in CRC microenvironment with its specific mechanism. METHODS: Firstly, the transcriptional expression profiles of CRC were downloaded from TCGA database and integrated via R software (4.1.2). Between CRC and normal colorectal tissues, ALKBH5 mRNA expressions were compared (Wilcoxon rank-sum). We further identified the expression levels of ALKBH5 in CRC tissues and cell lines through quantitative PCR, western blot, and immunohistochemistry. Then, how ALKBH5 affects the biological behaviors of CRC cells were confirmed by gain- and loss-of-function analysis. Furthermore, the relationship between ALKBH5 level and 22 tumor-infiltrating immune cells was examined through CIBERSORT in R software. Furthermore, we explored the correlation between ALKBH5 expression and tumor-infiltrated CD8+, CD4+ and regulatory T cells by utilizing the TIMER database. Finally, the association between chemokines and CD8+ T cells infiltration in CRC was analyzed using GEPIA online database. qRT-PCR, WB and IHC were used to further determine the effect of ALKBH5 on NF-κB-CCL5 signaling axis and CD8+ T cells infiltration. RESULTS: Clinically, ALKBH5 expression was downregulated in CRC and low levels of ALKBH5 expression were correlated with poor overall survival (OS). Functionally, overexpression of ALKBH5 reduced the proliferation, migration and invasion of CRC cells, and vice versa. Overexpression of ALKBH5 suppresses NF-κB pathway, thus reduces CCL5 expression and promotes CD8+ T cells infiltration in CRC microenvironment. CONCLUSIONS: ALKBH5 is poorly expressed in CRC, and overexpression of ALKBH5 attenuates CRC malignant progression by inhibiting CRC cell proliferation, migration, invasion and promoting CD8+ T cells infiltration in the tumor microenvironment through NF-κB-CCL5 axis.

12.
Pharmacol Res ; : 106787, 2023 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-37224894

RESUMO

Polyphenols, which are naturally occurring bioactive compounds in fruits and vegetables, are emerging as potential therapeutics for neurological disorders such as Parkinson's disease (PD). Polyphenols have diverse biological activities, such as anti-oxidative, anti-inflammatory, anti-apoptotic, and α-synuclein aggregation inhibitory effects, which could ameliorate PD pathogenesis. Studies have shown that polyphenols are capable of regulating the gut microbiota (GM) and its metabolites; in turn, polyphenols are extensively metabolized by the GM, resulting in the generation of bioactive secondary metabolites. These metabolites may regulate various physiological processes, including inflammatory responses, energy metabolism, intercellular communication, and host immunity. With increasing recognition of the importance of the microbiota-gut-brain axis (MGBA) in PD etiology, polyphenols have attracted growing attention as MGBA regulators. In order to address the potential therapeutic role of polyphenolic compounds in PD, we focused on MGBA. DATA AVAILABILITY: Data will be made available on request.

13.
Artigo em Inglês | MEDLINE | ID: mdl-37222253

RESUMO

OBJECTIVE: To assess the effects of estradiol (E2) on trigger day on cumulative live birth rates (CLBRs), and pregnancy outcomes after fresh and frozen-thawed embryo transfer (FET). METHODS: This multicenter retrospective cohort study included 42 315 patients from five reproductive centers. Six subgroups were divided according to E2 on trigger day (<1000, 1000-2000, 2000-3000, 3000-4000, 4000-5000, >5000 pg/mL). Smooth curve fitting and nonlinear mixed-effects models were used. RESULTS: When E2 was <5500 pg/mL, the CLBR increased by 10% for every 1000 pg/mL increase in E2. When E2 was between 5500 and 13 281 pg/mL, CLBR increased by 1.8% for every 1000 pg/mL increase in E2. When E2 was >13 281 pg/mL, CLBR decreased by 3% for every 1000 pg/mL increase in E2. From group E2 < 1000 to group E2 > 5000 pg/mL, pregnancy and live birth rates in fresh cycles were not related to E2. The live birth rate after FET was higher in the E2 ≥ 5000 pg/mL group than in the E2 < 1000 pg/mL group (odds ratio [OR] 4.03, and 95% confidence interval [CI] 3.74-4.35; adjusted OR 1.20, 95% CI 1.05-1.37). CONCLUSION: CLBR is associated with E2 on trigger day in a segmented manner. Pregnancy and live birth rates in fresh cycles were not associated with E2. The live birth rate in FET cycles was highest when E2 ≥ 5000 pg/mL.

14.
Mol Neurobiol ; 60(7): 4150-4160, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37171577

RESUMO

Cyclin F (CCNF) variants have been found to be associated with amyotrophic lateral sclerosis (ALS)/frontotemporal dementia (FTD). However, the genetic and clinical characteristics of ALS patients who carry CCNF variants are largely unknown. Genetic analysis was performed for 1587 Chinese ALS patients, and missense variants were predicted by software analyses. Additionally, we searched PubMed, Embase, and Web of Science for relevant literature and conducted a meta-analysis of the frequency of variants. In our ALS cohort, we identified 29 nonsynonymous variants in 41 ALS patients. Among these ALS patients, 18 (1.1%) were carriers of 15 rare missense variants that were considered probably pathogenic variants, and 11 of 15 variants were novel. Seven relevant studies were identified, and a total of 43 CCNF variants in 59 ALS patients with a frequency of 0.8% were reported. The ratio of males to females in our cohort (10/8) was similar to that in Caucasian populations (4/7) and significantly higher than that in Asian populations (10/1). The proportion of bulbar onset in Caucasian CCNF carriers was similar to our cohort (25.0 vs. 27.8%); however, bulbar onset had never been reported in previous Asian studies (0/11). FTD was not found in CCNF carriers in previous Asian studies and our cohort, but it has been reported in a FALS cohort (1/75) of Caucasian individuals. There were some differences in the clinical characteristics among different ethnic ALS populations. More basic scientific studies are needed to elucidate the pathogenic mechanisms and genotype-phenotype associations of CCNF variants.


Assuntos
Esclerose Amiotrófica Lateral , Demência Frontotemporal , Humanos , Masculino , Feminino , Esclerose Amiotrófica Lateral/genética , População do Leste Asiático , Demência Frontotemporal/genética , Estudos de Associação Genética , Mutação de Sentido Incorreto/genética , Mutação , Ciclinas
15.
Foods ; 12(10)2023 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-37238834

RESUMO

The selection of base liquor plays a crucial role in the flavor of soaked greengage wine. This study aimed to investigate the effects of different base liquor treatments on the physicochemical characteristics and aroma composition of greengage wine. We carried out a comprehensive analysis using HPLC for the determination of organic acids and GC-MS for the determination of volatile aroma compounds, combined with sensory evaluation. The results showed that the red and yellow colors were the darkest in the high-alcohol group, while the citric acid content was the highest in the sake group (21.95 ± 2.19 g/L). In addition, the greengage wine steeped in 50% edible alcohol had more terpenes, a significantly higher concentration of acid-lipid compounds, and a more intense aroma compared to that of the low-alcohol group, whose typical aroma compounds were greatly reduced. The sensory results showed that the greengage wine treated with baijiu had a distinct alcoholic flavor, while almond flavors were more intense in the greengage wine treated with 15% edible alcohol. In this study, base liquor was used as the main influencing factor to provide new research ideas for the flavor optimization of soaked greengage wine.

17.
Int J Oncol ; 63(1)2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37232381

RESUMO

Following the publication of this paper, it was drawn to the Editor's attention by a concerned reader that several of the protein bands featured in the western blot assay data shown in Fig. 3A­D on p. 2147 were strikingly similar to other protein bands, both comparing the data within the same gel slices and comparing the data across the four different parts of the figure. In addition, the control blots featured in Fig. 3A, B and D had already appeared in a different form written by (largely) different authors at different research institutes. After having conducted an independent review of the data in this Figure in the Editorial Office, the concerns of the reader were found to be validated. Therefore, since contentious data in the above article had already been published prior to its submission to International Journal of Oncology, and owing to an overall lack of confidence in the presented data, the Editor has decided that this paper should be retracted from the Journal. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a reply. The Editor apologizes to the readership for any inconvenience caused. [International Journal of Oncology 45: 2143­2152, 2014; DOI: 10.3892/ijo.2014.2596].

18.
Food Chem ; 424: 136349, 2023 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-37244185

RESUMO

Acrylamide (AA) and heterocyclic aromatic amines (HAAs), as classic hazards produced during food thermal processing, have been widely concerned, but because of their polarity difference, it is very difficult to detect these contaminants simultaneously. Herein, novel cysteine (Cys)-functionalized magnetic covalent organic frameworks (Fe3O4@COF@Cys) were synthesized via a thiol-ene click strategy and then used as adsorbents for magnetic solid-phase extraction (MSPE). Benefiting from the hydrophobic properties of COFs and the modification of hydrophilic Cys, AA and HAAs could be enriched simultaneously. Then, a rapid and reliable method based on MSPE coupled with HPLC-MS/MS was developed for the simultaneous detection of AA and 5 HAAs in thermally processed foods. The proposed method showed good linearity (R2 ≥ 0.9987) with satisfactory limits of detection (0.012-0.210 µg kg-1) and recoveries (90.4-102.8%). Actual sample analysis showed that the levels of AA and HAAs in French fries were affected by frying time and temperature, water activity of samples, content and type of reaction precursors, and reuse of oils.

19.
Sci Total Environ ; : 164440, 2023 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-37244608

RESUMO

Cyanobacteria and their metabolites are one of the primary precursors of disinfection by-products (DBPs) in natural water environments. However, few studies have investigated whether the production of DBPs by cyanobacteria changes under complex environmental conditions and possible mechanisms underlying these changes. Therefore, we investigated the effects of algal growth phase, water temperature, pH, illumination and nutrients on the production of trihalomethane formation potential (THMFPs) by Microcystis aeruginosa in four algal metabolic fractions, that is, hydrophilic extracellular organic matter (HPI-EOM), hydrophobic EOM (HPO-EOM), hydrophilic intracellular organic matter (HPI-IOM) and hydrophobic IOM (HPO-IOM). Additionally, correlations between THMFPs and some typical algal metabolite surrogates were analyzed. The results showed that the productivity of THMFPs by M. aeruginosa in EOM could be affected significantly by the algal growth phase and incubation conditions, while the IOM productivity varied insignificantly. M. aeruginosa in the death phase could secrete more EOM and have a higher THMFP productivity than those in the exponential or stationary phases. Cyanobacteria grown under harsh conditions could have increased THMFP productivity in EOM by increasing the reactivity of algal metabolites with chlorine, for example, under low pH conditions, and secreting more metabolites in EOM, for example, under low temperature or nutrient limitation conditions. Polysaccharides were responsible for the enhanced THMFP productivity in HPI-EOM fraction, and a significant linear correlation was found between the concentration of polysaccharides and THMFPs (r = 0.8307). However, THMFPs in HPO-EOM did not correlate with dissolved organic carbon (DOC), ultraviolet absorbance at 254 nm (UV254), specific UV absorbance (SUVA) and cell density. Thus, we could not specify the kind of algal metabolites that contribute to the increased THMFPs in the HPO-EOM fraction under harsh growth conditions. Compared with the case in EOM, the THMFPs in IOM were more stable and correlated with the cell density and total amount of IOM. The results implied that the THMFPs in the EOM were sensitive to growth conditions and were independent of algal density. Considering the fact that traditional water treatment plants cannot remove dissolved organics as efficiently as algal cells, the increased THMFP productivity in EOM by M. aeruginosa under harsh growth conditions could be a potentially serious threat to the safety of the water supply.

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