R = 0.45 and R = 0.58, respectively). These findings suggest that specific inflammatory response and/or regulation locally occurs within the lower airways.Few data on cytokine profiles in bronchoalveolar lavage fluid (BALF) are available for racehorses with mild/moderate equine asthma (EA); cytological diagnosis being most frequently made from only one lung. The purpose of the study was to compare cytokine mRNA expressions and protein concentrations in BALF from both lungs. As part of a larger study, 250 ml saline was randomly instilled in one lung and 500 ml in the contralateral lung of 30 clinically healthy Standardbred racehorses. This procedure was repeated 72 h later, inversing the volume per lung. Cytological cut-off values for diagnosis of mild EA was neutrophil proportions > 10% when instilling 250 ml. Eleven horses that exhibited unilateral mild inflammatory cytology  were enrolled. Protein concentrations were not significantly different between lungs, for any of the investigated cytokines. Relative mRNA expression of IL-1β (3.887 ± 0.929) and IL-10 (3.225 ± 0.516) were significantly higher in BALF from mild inflammatory lungs when compared with non-inflammatory ones ; and also significantly correlated with neutrophil proportions ( Based on the multiple similarities shared with asthma in humans, equine asthma (EA) is a translational terminology which has recently been suggested to describe horses of any age with chronic, reversible and mainly environment related lower airway inflammation , 2. ThisThe pathophysiology of mild/moderate EA, and subsequent corresponding endotypes, remains poorly understood. Elucidating local and systemic immune responses associated with EA has, furthermore, been recommended as one research priority by the recently revised Consensus Statement on IAD . Very feEither RT-PCR or ELISA has been performed previously, in order to determine BALF cytokine profiles of horses with mild/moderate EA. To date, mRNA expression for up to 10 different cytokines have been documented –9; proteTo date, the possibility of a local and specific immune response in horses with mild EA still remains to be considered. We hypothesized that BALF cytological discrepancy between lungs within the same horse would be associated with specific BALF cytokine profiles, especially with regard to the T-helper (Th) polarization. The aim of this study was to investigate the pattern of mRNA expression and corresponding protein concentration for a large panel of cytokines, in BALF harvested from both lungs of horses with unilateral mild inflammatory cytology.A total of 30 client-owned Standardbred racehorses (aged 3–9 years), located in three different training centers, were randomly included in this prospective observational study. Horses had no recent history of coughing or nasal discharge and were free of any clinical evidence of respiratory disease when examined at rest, as described previously . The stuBALF samples from dorsal lung lobes were systematically collected twice at 72 h interval; both in the left and right lungs by the same operator, according to the methodology previously described . For eacOnly BALF samples originating from instillations of 250 ml were investigated in the current study. One aliquot for each sample (4 ml) was kept refrigerated (+4°C) in EDTA tubes until cytological preparation within 12 h. For cytokine investigations, 50 ml of BALF were immediately centrifuged  and cell pellets dissociated in RNAprotect Cell Reagent . Both cells and supernatants were kept frozen (−80°C) until further used.BALF total cell count was performed automatically . Differential cell count was performed on 300 leukocytes, after cytocentrifugation of 300 μl BALF  and May-Grünwald-Giemsa staining . Any BALHorses were then classified in three groups, based on BALF status from each lung. Each horse exhibiting two “non-inflammatory” BALF samples was classified as “CTL (-)”; each horse exhibiting two “mild inflammatory” BALF samples was classified as “CTL (+)”; each horse exhibiting one “non-inflammatory” and one “mild inflammatory” BALF samples was classified as “Mixed.” For comparisons within groups, paired BALF samples for each horse were further categorized as “high” and “low” according to the corresponding neutrophil differential count, irrespectively to the lung side of origin.Total RNA was extracted from cell pellets using RNeasy Plus Mini Kit  and RNA quality assessed using NanoDrop 2000c Spectrophotometer . cDNA were generated using Superscript First Strand System for RT-PCR , and stored at −20°C until further use. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was previously found to be the most stably expressed housekeeping gene  in BAL oLung2-(cytokine gene Ct- GAPDH Ct) Lung1; lung 1 and lung 2, respectively, correspond to BALF samples with the lowest and highest neutrophil proportions for the same horse, and Ct is the crossing point above background fluorescence. Variation in cytokine gene expression for each horse was calculated with GenEx6 .Changes in cytokine gene expression were calculated by relative quantitation using the ΔΔCt method , where ΔAfter concentrating the BALF 10-fold , IFN-γ, IL-1β, IL-4, IL-8, IL-10, IL-17, and TNF-α concentrations were measured in duplicates by ELISA, according to the manufacturer's instructions with minor modifications . Concentpost-hoc test, with sampling day and training center as covariates. Correlations between neutrophil proportions and cytokines mRNA expression were determined by Pearson's correlation coefficient. Neutrophil proportions and cytokine concentrations were expressed as median (1st−3rd quartile); cytokine gene expressions per paired samples were expressed as mean fold change ± SEM. Analyses were conducted using NCSS9  and Prism 7 .Continuous data which were not normally distributed, as assessed by Shapiro-Wilk W test, were log 10 transformed. Within each group of horses, fold-changes in cytokine gene expression and protein concentrations were evaluated by ANOVA  and Tukey-Kramer's P < 0.001) but not in any of the “CTL” groups ,” 12 horses as “CTL (+)” and 11 were classified as “Mixed,” based on BALF neutrophil proportions . Within ” groups .P = 0.008 and P = 0.01, respectively), and IL-4 mRNA expression within both “Mixed” and “CTL (+)” groups was significantly influenced by the day of sampling . Within the “Mixed” group (P = 0.003) and IL-10  were significantly higher in “mild inflammatory” BALF samples, when compared with “non-inflammatory” samples from the contralateral lung , controlling for confounding factors. Protein concentrations in BALF for either IL-1β or IL-10 were not significantly different among between lungs  and IL-10 , but not with their corresponding protein concentrations. Relative mRNA expressions of IFN-γ, IL-12, IL-4, IL-17, IL-23, IL6, IL-8, and TNF-α, as well as protein concentrations of IFN-γ, IL-4, IL-17, IL-8, and TNF-α in BALF of horses from the “Mixed” group were not significantly different between “mild inflammatory” and “non-inflammatory” samples  neutrophilic inflammation induced by the procedure , we deciThe higher relative mRNA expression of IL-1β in the “mild inflammatory” BALF is in agreement with previous studies for which neutrophilic mild/moderate EA subgroups were compared with controls , 9. ThisSimilarly to IL-6 and IL-8, relative mRNA expression of IL-17 was not significantly different between “mild inflammatory” and “non-inflammatory” samples. Overexpression of IL-17 has previously been documented for mild/moderate EA in one study only , 29. RelAlike previous studies on horses suffering from mild/moderate EA , 8, relaThis study suggests that immune microenvironment could differ from one lung to the other one in the same horse with unilateral airway neutrophilia. The results reported here support, for mRNA expression but not for protein concentration, previous cytological evidences highlighting a specific but localized and asymmetrical inflammatory response. This study however provides no evidence for a Th polarization associated with mild neutrophilic airway inflammation. Further investigations are needed concerning the unbalanced pro- and anti-inflammatory cytokines pattern in the airways of horses with EA.All datasets generated for this study are included in the article/The animal study was reviewed and approved by Comité d'Éthique en Expérimentation Animale Pays de la Loire. Written informed consent was obtained from the owners for the participation of their animals in this study.ER conceived and designed the study. EH, MO, M-PT, MD, AC, and ER contributed to data collection and analysis. EH, MO, SP, RP, and ER interpreted the data. EH, RP, and ER wrote the manuscript. All authors read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Platinum resistance is an important cause of clinical recurrence and death for ovarian cancer. This study tries to systematically explore the molecular mechanisms for platinum resistance in ovarian cancer and identify regulatory genes and pathways via text mining and other methods.Genes in abstracts of associated literatures were identified. Gene ontology and protein-protein interaction (PPI) network analysis were performed. Then co-occurrence between genes and ovarian cancer subtypes were carried out followed by cluster analysis.Genes with highest frequencies are mostly involved in DNA repair, apoptosis, metal transport and drug detoxification, which are closely related to platinum resistance. Gene ontology analysis confirms this result. Some proteins such as TP53, HSP90, ESR1, AKT1, BRCA1, EGFR and CTNNB1 work as hub nodes in PPI network. According to cluster analysis, specific genes were highlighted in each subtype of ovarian cancer, indicating that various subtypes may have different resistance mechanisms respectively.Platinum resistance in ovarian cancer involves complicated signaling pathways and different subtypes may have specific mechanisms. Text mining, combined with other bio-information methods, is an effective way for systematic analysis. Ovarian cancer is the most lethal cause of all gynecological malignancies . Due to Although a plenty of genes and pathways have been investigated for platinum resistance in ovarian cancer, mechanisms of drug resistance are still not fully understood. Most researchers examined only a small part of genes, meanwhile the majority of them focused on specific subtypes of ovarian cancer. As platinum resistance seems to be regulated by sophisticate molecular networks, we try to systematically assess reported genes with text mining and other bioinformatics methods, quantitatively describe their relationships and make prediction of potential regulatory molecules and pathways in this study.Ovarian cancer AND (cisplatin OR carboplatin) were used as retrieval statement on Pubmed and 6160 literatures were listed . All abstracts were collected from PubMed retrieval system. Genes and proteins were identified with ABNER (V1.5) [“platinum”, “platin”, “cisplatin”, “DDP”, “carboplatin” and “CBP” were selected. Similarly, both “resistance” and “resistant” were identified. Only the genes that co-appeared with these two groups of words in the same sentence will be treated. If a gene appeared several times in one sentence, it would be counted once. Word frequency analysis was performed with Microsoft Excel 2010. Gene ontology analysis was carried with FunRich (V3.0) software [p-value were corrected with Bonferroni method.The methods for data preparation and gene identification have been described previously . BrieflyR (V1.5) , 9 and w“serous”, “mucinous”, “endometrioid”, “clear cell cancer” or “OCCC”) using HemI (V1.0) [Protein-protein interaction (PPI) network analysis was performed using Cytoscape (V3.4.0). Plugins such as BisoGenet  and CytoI (V1.0)  with maxTP53 were mentioned more than 100 times, while ABCB1, AKT1, ERCC1 and other genes were also widely studied in the past years.According to the criterion of frequency analysis, 473 genes were identified within 6160 abstracts and top genes among them (count≥15) were listed in Table p < 0.05) in platinum resistance were shown in Table To explore the functions of these genes, gene ontology (GO) analysis was carried out. Significant biological processes that may involve  software and its plugins. The interactions were illustrated in Fig. .4.0 softBased on histopathology, ovarian cancer can be mainly classified into four subtypes: serous, mucinous, endometrioid and ovarian cancer of clear cell (OCCC) . Each maserous”, “mucinous”, “endometrioid” and “clear cell” (or OCCC) were collected respectively, then cluster analysis were performed. As shown in Fig. TP53 are commonly focused in most subtypes. By comparison, BCL2 and AKT1 were frequently mentioned in endometrioid cancer while ERBB2 and AGR3 were repeatedly mentioned in mucinous cancer. Such genes may be regarded as specific regulators or markers for each subtype.To investigate the specific regulatory molecules for each subtype, genes co-appearing with “Cisplatin and carboplatin exert antitumor effects by binding to DNA and forming cross-links, thus disrupts DNA structure and finally results in cell apoptosis . DysreguBCL2 can inhibit cell death induced by cytotoxic factors such as chemotherapeutic drugs and enhance cell resistance [According to Table sistance , 31.ABCB1 (MDR1) and ABCC1 (MRP1) belong to ATP binding cassette (ABC) transport protein family, which works as ATP-dependent drug efflux pump and is responsible for decreased platinum accumulation [ABCG2 (count = 13) and ABCC2 (count = 10) have similar functions though not listed in Table SLC31A1 [CTR1), a member of copper transporter family, which plays a significant role in platinum uptake [For platinum accumulation, both mulation , 33. Amo SLC31A1   is a critical member of nucleotide excision repair induced by platinum [BRCA1 [BRCA2 [PARP1 can recognize DNA lesions and modifies various nuclear proteins which are involved in the regulation of DNA repair [For DNA damage/repair, platinum . Meanwhim [BRCA1  and BRCA1 [BRCA2  exert thA repair .GSTA1 (count = 12) and GSTP1 (count = 9) belong to the top 10% of all identified genes though not listed in Table Both AKT1, EGFR, PIK3CA, MAPK1, NFKB1 and MTOR, are difficult to be classified. All of them have multiple functions in physiological and pathological processes and are regarded as key nodes in platinum-resistance signaling network , but the rate is generally low in clear cell carcinomas [XPB and ERCC1), were found to be preferentially expressed in ovarian clear cell carcinomas [There are specific genomic alterations and gene-expression patterns for different subtypes of ovarian cancer. According to previous reports, rcinomas , 47. Mearcinomas , 49. ThaA potential limitation in this study is the performance of text mining. It can recognize names of genes and proteins, calculate their frequencies and judge the functions of them via co-occurrence analysis, but it cannot really “understand” literatures. However, it is still an effective method to quantitatively assess gene functions and their relationships, especially for comprehensive analysis with large input data.


Scientific Reports 10.1038/srep21848, published online 22 February 2016Correction to: Pongo pygmaeus. In fact, the studied individual was a Sumatran orangutan Pongo abelii.This Article contains an error. Among the studied species, the orangutan was erroneously specified as Bornean orangutan In the Results section,Pongo pygmaeus) retina .”“The same held true for the hominid orangutan (should read:Pongo abelii) retina .”“The same held true for the hominid orangutan (The error is also present in Figure 1, the correct Figure Finally, the error is present in Table S1, wherePongo pygmaeus”“Bornean orangutan, should read:, Pongo abelii”“Sumatran orangutanThis error does not alter the conclusions of the paper.


However, the in vivo rate of this condensation reaction is unknown and its possible contribution to growth remains elusive. Here, we used microbial platforms to assess the rate of this condensation in the cellular environment. We constructed Escherichia coli strains lacking the enzymes that naturally produce 5,10-methylene-THF. These strains were able to grow on minimal medium only when equipped with a sarcosine (N-methyl-glycine) oxidation pathway that sustained a high cellular concentration of formaldehyde, which spontaneously reacts with THF to produce 5,10-methylene-THF. We used flux balance analysis to derive the rate of the spontaneous condensation from the observed growth rate. According to this, we calculated that a microorganism obtaining its entire biomass via the spontaneous condensation of formaldehyde with THF would have a doubling time of more than three weeks. Hence, this spontaneous reaction is unlikely to serve as an effective route for formaldehyde assimilation.Formaldehyde is a highly reactive compound that participates in multiple spontaneous reactions, but these are mostly deleterious and damage cellular components. In contrast, the spontaneous condensation of formaldehyde with tetrahydrofolate (THF) has been proposed to contribute to the assimilation of this intermediate during growth on C Most metabolic conversions are catalyzed by dedicated enzymes or by promiscuous enzymes that evolved to catalyze other reactions . Neverth1 feedstock into multi-carbon metabolites. Some of the formaldehyde condensation reactions in methylotrophic organisms were speculated to be spontaneous. For example, condensation of formaldehyde with tetrahydromethanopterin, which plays a key role in the methylotrophic serine cycle  should be lower than [THF] + , and the reverse reaction  further lowers the net formaldehyde condensation flux, the in vivo condensation flux seems likely to be 2–3 orders of magnitude higher than that expected from the in vitro parameters. While the intracellular concentration of formaldehyde might be somewhat higher than that measured in the medium, it is highly unlikely that this discrepancy accounts for the 2–3 orders of magnitude difference. Hence, it is rather clear that under cellular conditions, the condensation rate of formaldehyde with THF is substantially higher than that measured in simple model in vitro systems, further emphasizing the importance of the analysis performed here.How does the in vivo derived condensation flux compare with that expected using the in vitro measured parameters? Using the previously measured kinetic parameters, the rate of 5,10-methylene-THF formation equals  s−1 M−2 . Even ifE. coli the formaldehyde-THF condensation reaction as well as the specific reactions of the serine cycle . Again, we calculated the phenotypic phase plane of the formaldehyde condensation reaction, assuming formaldehyde to be the sole carbon source. As expected, the correlation factor between the growth rate and the formaldehyde condensation rate was much higher than before, >20 mmol gCDW−1 : as 2.3% of the carbons in biomass are derived from non-thymidine C1 moieties [−1, very close to the value derived by the computational modeling.) This growth rate is equivalent to a doubling time of 580 h, i.e., 24 days. Obviously, such a growth rate is impractical for most microorganisms, ruling out the spontaneous condensation of formaldehyde and THF as an effective route for formaldehyde assimilation.Next, we asked what would be the growth rate of an organism growing on formaldehyde (or a precursor thereof) using the serine cycle and assuming that 5,10-methylene-THF is produced solely via the spontaneous condensation of formaldehyde with THF. To assess this rate, we added to the model of moieties , the exp2-dependent methanol oxidase, both strongly favoring methanol oxidation, is expected to enable growth with methanol just as SOX enabled growth with sarcosine. The strict necessity for thymidine supplementation could be attributed to the poor kinetics of the enzyme that generates it from 5,10-methylene-THF. This kinetic constraint does not seem to hamper thymidine biosynthesis in wild-type E. coli presumably because the biosynthesis of 5,10-methylene-THF is not limiting. In our strain, the low production rate of 5,10-methylene-THF likely generates a “competition” between its various metabolic sinks  sarcosine oxidation sustained a high concentration of formaldehyde and (ii) thymidine was added to the medium. The failure to obtain growth with methanol as a source of formaldehyde might be attributed to the use of an NAD-dependent MDH, which strongly favors formaldehyde reduction over methanol oxidation. A quinone-dependent MDH or an Oic sinks , such thE. coli strains established here could be cultivated in a turbidostat regime [As we demonstrated, a microorganism obtaining its entire biomass via the spontaneous condensation of formaldehyde with THF would be constrained to a very low growth rate, having a doubling time of more than three weeks. However, it is important to remember that the spontaneous condensation rate might differ between organisms. For example, it could be improved by increasing the cellular concentration of THF or operating at a higher formaldehyde concentration. Moreover, changing the cellular conditions—temperature, pH, ionic strength, and concentrations of different ions—might alter the rate at which formaldehyde reacts with THF. Whether such changes could substantially increase the spontaneous production rate of 5,10-methylene-THF, thus enabling a more reasonable growth rate, remains to be seen. It will be interesting to use adaptive laboratory evolution  to try tt regime  selectint regime  has evolDespite the kinetic barrier, assimilation of formaldehyde via its spontaneous condensation with THF might be advantageous in conditions that favor high biomass yield rather than growth rate. This is due to two reasons. First, microorganisms that grow on methanol via the serine cycle use a long, tetrahydromethanopterin-dependent route . BypassiE. coli K-12 MG1655 derived strain SIJ488 [Corynebacterium glutamicum R and sarcosine oxidase (SOX) from Bacillus sp. B-0618 were overexpressed in a pZASS plasmid, which has a p15A origin, a constitutive promoter pgi-20, and a streptomycin selection marker; gene sequences and cloning details were provided previously in ref. [All strains used in this study are listed in n SIJ488  was usedn SIJ488  and antin SIJ488 . Colonie in ref. .2HPO4, 22 mM KH2PO4, 8.6 mM NaCl, 18.7 mM NH4Cl, 2 mM MgSO4, and 100 μM CaCl2), supplemented with trace elements . Additional 50 μM MnCl2 was added to relieve possible oxidative stress from sarcosine oxidation [1 supplements were added according to the specific experiment: 0.3 mM of thymidine, 0.3 mM of inosine, 0.3 mM of methionine, and 6 µM of pantothenate [LB medium  was used for genomic strain modifications. Antibiotics were used at the following concentrations: kanamycin, 50 μg/mL and streptomycin, 100 μg/mL. Growth experiments were performed in M9 minimal media . Cell densities were measured by photometer  at 600 nm. Cell densities at exponential phase were fitted to 0N and N are cell densities at start and at time t, and μ is growth rate. Student’s t-test was used for the statistical analysis.Strains were precultured in 4 mL M9 minimal medium with all Cg, 4 °C. Formaldehyde concentration in culture supernatants was determined by Nash assay [At the indicated time points, 140 μL of culture were collected and centrifuged for 10 min at 3300× sh assay . 125 μL 13C labeled sarcosine . Experiments were performed in duplicate. Cells were harvested at the late exponential phase (OD 0.5). The 2 mL of culture was harvested and washed by centrifugation. Protein biomass was hydrolyzed with 6 M HCl, at 95 °C for 24 h [2 at 95 °C.For stationary isotope tracing of proteinogenic amino acids, C1AΔ3 strain was grown in M9 with 10 mM of glucose, 0.3 mM of thymidine, and 1 mM of either unlabeled sarcosine or methyl-for 24 h . The sam18 reversed phase column . Of formic acid 0.1% in H2O (A) and 0.1% of formic acid in acetonitrile (B) were the mobile phases. The flow rate was 0.4 mL/min and the gradient was 0–1 min—99% A; 1–5 min—linear gradient from 99% A to 82%; 5–6 min—linear gradient from 82% A to 1% A; 6–8 min—kept at 1% A; 8–8.5 min—linear gradient to 99% A; and 8.5–11 min—re-equilibrate. Mass spectra were acquired using an Exactive mass spectrometer  in positive ionization mode, with a scan range of 50.0–300.0 m/z. The spectra were recorded during the first 5 min of the LC gradients. Data analysis was performed using Xcalibur . Determination of retention times was performed by analyzing amino-acid standards (Sigma-Aldrich) under the same conditions.Hydrolyzed amino acids were analyzed with ultra-performance liquid chromatography coupled with electrospray mass spectrometry (UPLC–ESI–MS) as previously described . Chromathttps://gitlab.com/hi-he/wenzhangfujian under the directory of “2020_formaldehyde condensation”.For Flux Balance Analysis and phenotypic phase plane calculations, we used the COBRApy package . All scriML1515 model [E. coli cells.Reactions POR5 and GLYCK were removed from the model since they are not present in The stoichiometry of the reaction THD2pp was changed, as the translocation uses only one proton instead of two.PFL and OBTFL were removed, since they are catalyzed by oxygen sensitive enzymes and therefore not active in aerobic conditions.Based on gathered evidence, FTHFLi was removed. This reaction is especially important in our case, since it could allow charging of THF with formate directly.We changed the directionality of three reactions in the nucleotide biosynthesis pathway  to be irreversible, in order to avoid nucleotides degradation to charge THF, since that is very unlikely to occur.We changed the thymidine periplasmic symporter reaction (THYMt3pp) to be reversible, based on our experimental evidence (adding thymidine to the media rescued growth in certain conditions).We knocked out the glycine cleavage system reaction and the GlyA reaction (GLYCL and GHMT2r).We added the formaldehyde-THF spontaneous condensation (THFSPONT) reaction: thf_c + fald_c -> mlthf_c + h2o_c.Adjustments made to 15 model :ReactionRemoved the following reactions to avoid potential carbon fixation cycles: GLXCL, GLYCL, THRD, THRA2, THRA, SERD_L.Added NAD-dependent formate dehydrogenase (FDH), enabling the utilization of the formate that is created from formaldehyde as source of reducing power.Added the crotonoyl-CoA carboxylase reaction (EtMaCoA), required for the ethylmalonyl-CoA cycle: b2coa_c + nadph_c + co2_c + q8_c + h2o_c -> glx_c + ppcoa_c + nadp_c + q8h2_c.Added the propionyl-CoA carboxylase reaction (MMCDr), as required for recycling the propionyl-CoA, which was created by EtMaCoA: ppcoa_c + atp_c + hco3_c -> mmcoa__S_c + adp_c + h_c + pi_c.Made the methylmalonyl-CoA mutase reaction (MMM) reversible so that the methylmalonyl-CoA (mmcoa__S_c) can be recycled.○serine-hydroxypyruvate amino-transferase (SGAT1): ser__L_c + akg_c -> glu__L_c + hpyr_c.○glycine-glyoxylate amino-transferase (SGAT2): gly_c + akg_c -> glx_c + glu__L_c.○malate thiokinase (MTK): mal__L_c + atp_c + coa_c -> malylcoa_c + adp_c + pi_c.○malyl-CoA lyase (MCL): malylcoa_c -> accoa_c + glx_c.Added the reactions of the serine cycle:We made further adjustments to the model to enable growth on formaldehyde via the serine cycle and ethylmalonyl-CoA cycle:g, 10 min, 4 °C) and resuspended in 10 mL of 50 mM HEPES-CHES (2-(N-cyclohexylamino)ethanesulfonic acid; pH 7.8 ± 0.05) containing 2% (w/v) ascorbic acid and 10 mM β-mercaptoethanol (extraction buffer). The cells were disrupted using a FastPrep Instrument , boiled for 10 min, and centrifuged . The supernatants were collected and treated with AtGGH2, as described by Ramos-Parra et al. [2HPO4 and 59 mM H3PO4) and B (75% phase A and 25% acetonitrile). Detection was achieved with an electrochemical cell adjusted to 100, 200, 300, and 400 mV, and quantification was performed using commercial standards .75 mL of each culture was incubated in 250 mL Erlenmeyer flask. At the late exponential phase, cells of each strain equivalent to approximately 5 mg total protein  were hara et al. . Folatesa et al. . Separat


Pseudomonas aeruginosa”“Gene regulatory network inference and analysis of multidrug-resistant , DOI number: 10.1590/0074-02760190105, published in Mem Inst Oswaldo Cruz, Rio de Janeiro, Vol. 114: e190105, 2019: In the article On page 7, Table I, where it reads:“PA4851_19380” It should read: “AL347_05155”On page 7, where it reads:np20 and PA4851_19380 (homologous to PA1520).” “After pinpointing the hubs, an analysis was performed to determine whether they are interconnected (through direct or indirect interactions). Only two hubs were found to not be interconnected, It should read: np20 and AL347_05155 (homologous to PA1520).”“After pinpointing the hubs, an analysis was performed to determine whether they are interconnected (through direct or indirect interactions). Only two hubs were found to not be interconnected, On page 10, where it reads:np20) and purine metabolism (PA4851_19380), which are fundamental to bacterial survival, but can be considered somewhat independent of other processes, and are only triggered under specific conditions.” “In fact, they are related to processes such as zinc uptake (It should read: np20) and purine metabolism (AL347_05155), which are fundamental to bacterial survival, but can be considered somewhat independent of other processes, and are only triggered under specific conditions.”“In fact, they are related to processes such as zinc uptake  One clear trend is that the CCBH4851 GRN represents a substantial improvement in terms of network completeness, since is includes more nodes, edges and network motifs.” It should read: “Table II shows a comparison of network statistics between the CCBH4851 GRN and the GRN network published by Galán-Vásquez et al. (12) One clear trend is that the CCBH4851 GRN represents a substantial improvement in terms of network completeness, since it includes more nodes, edges and network motifs.”On page 4, On page 8, The Supplementary material were corrected and attached in the link below: https://memorias.ioc.fiocruz.br/images/revistas/2020/115/0105er_sd.xlsx


The present study aimed to determine whether whole-body fat oxidation and muscle deoxygenation kinetics parameters during exercise were related in individuals with different aerobic fitness levels.–1⋅min–1] and 11 active individuals (–1⋅min–1) performed a maximal incremental cycling test to determine max) eliciting maximal fat oxidation (MFO). The muscle deoxygenation response was fitted with a double linear model. The slope of the first parts of the kinetics (a1) and the breakpoint ([HHb]BP) were determined.Eleven cyclists BP occurred at a higher absolute intensity (p < 0.001) than in active individuals. max, and [HHb]BP . MFO and Fatmax were both correlated with [HHb]BP  and tended to be negatively correlated with a1 .MFO (2 delivery and consumption supporting higher fat oxidation rates.This study showed that whole-body fat oxidation and muscle deoxygenation kinetics were both related to aerobic fitness and that a relationship between the two kinetics exists. Individuals with greater aerobic fitness may have a delayed reliance on glycolytic metabolism at higher exercise intensities because of a longer maintained balance between O Regular endurance training leads to development of the muscle capillary network and marked changes in mitochondrial content and activity or in muscle fiber type composition see . This lemax”  to mathematically characterize the shape of the whole-body fat oxidation kinetics as a function of exercise intensity (see dilatation) projection of the fat oxidation kinetics compared to less-trained individuals.The whole-body fat oxidation rate measured during the incremental exercise test allows the estimation of an individual’s ability to oxidize fat . Fat oximax” . Achten ined men . In addisity see  may resu2) delivery and utilization into mitochondria may partly explain the greater fat oxidation capacity in endurance-trained athletes. In fact, endurance training leads to improved muscle blood perfusion. The increased muscular blood flow , followed by a near-plateau. Although somewhat controversial . Since endurance athletes have good oxidative capacity, their [HHb]BP may appear at higher absolute (A better oxygen (O(V.O2m) . Given tcapacity . This rarogation . Some auncreases . Howeverncreases  because oversial , [HHb]BPabsolute  and relaabsolute , reflectabsolute .BP, as well as upward and more dilated (greater dilatation) fat oxidation kinetics and (ii) a direct relationship between both kinetics would exist, with shallower muscular deoxygenation kinetics corresponding to higher fat oxidation rates.The adaptations to endurance training affecting aerobic fitness seem to influence both muscular deoxygenation and fat oxidation kinetics. However, to the best of our knowledge, the relationship between the two kinetics has never been investigated in healthy individuals of various aerobic fitness levels. Therefore, the purpose of the present study was to determine whether whole-body fat oxidation and muscle deoxygenation kinetics parameters during exercise were related in individuals with different aerobic fitness levels . It was hypothesized that (i) cyclists would have shallower muscular deoxygenation kinetics with a right-shifted [HHb]–1⋅min–1) and 11 non-specifically trained but active individuals] aged between 20 and 39 years participated in this study  were taken. Each subject then performed a maximal incremental exercise test on a cycle ergometer. For the remaining two sessions, the subjects performed two identical submaximal incremental exercise tests . Sessions 2 and 3 were identical and separated by a minimum of 7 days and a maximum of 10 days. The pre-experimental conditions were controlled to standardize measurements and to minimize inter- and intra-individual variations. Both tests were performed in the morning after a minimum 10-h overnight fast, after 3 days of a balanced diet , with no caffeine and no alcohol during the last 48 h, and the volunteers were asked to refrain from vigorous exercise during the day before the experimental trials. Before the beginning of the testing session, all the subjects confirmed that they had followed the nutritional indications, and the food diary was checked by the experimenters.Height, body mass, and composition were assessed. Body composition  was estimated from the Durnin and Womersley’s skinfold-thickness measurement method at four sites .2, 5.02% CO2); and (3) volume turbine calibration with a 3-L syringe. –1⋅min–1 with an increase of workload during the latter stages of the test) was considered as the MAP. The VT and RCP were estimated by three blinded and independent investigators: ventilatory equivalent method was used to determine both thresholds  corresponding to ∼3.14 and a constant of intensity named K, which corresponds to (π/100):Both submaximal experimental trials served to determine whole-body fat oxidation kinetics. Resting values were measured during a 10-min seated-rest. This was followed by a 10-min warm-up at 20% MAP. The workload was then increased by 7.5% MAP for the active group and 10% MAP for cyclists every 5 min until the end of the stage during which RER was 1.0 . For theorimetry . Fat oxiorimetry . The reswas used . This modilatation refers to the degree of dilation or retraction of the curve, symmetry modifies the symmetry of the standard basic sine curve, and translation refers to the right-left shift of the whole curve toward the abscissa axis (lipid) based on the SIN model and the calculated values of Elipid, as described in a previous study , and total hemoglobin (Hbtot) concentrations were continuously measured in the right vastus lateralis (VL) muscle using NIRS  at 50 Hz to determine muscle deoxygenation. The NIRS probe contains a three-diode emitter operating at wavelengths of 764 and 860 nm and a detector positioned 40 mm away from the emitter. After calibration of the device, the probe was positioned on shaved and cleaned skin over the right VL muscle, halfway between the great trochanter and lateral femoral condyle, parallel to the major axis of the thigh. The VL muscle was chosen to assess exercise-induced changes in [HHb] concentrations since this muscle is highly active during cycling , oxyhemoglobin (O cycling . After t cycling . A diffe cycling . Changesar model  as folloa1 is the slope of the linear regression before the breaking point ([HHb]BP); b is the y-intercept of the linear regression before the [HHb]BP; and a2 is the change in the slope of the linear regression after the [HHb]BP. The model parameters were estimated using an iterative procedure by minimizing the SMS of the differences between the estimated %Δ[HHb] based on the double linear model and the measured values of %Δ[HHb]. The slope of the linear regression after the [HHb]BP (a3) was then calculated as the sum of a1 and a2.where t-test or the Mann–Whitney test for non-parametric data. Independent correlations between the parameters of whole-body fat oxidation kinetics and those of the double linear model of %Δ[HHb] were performed by using Pearson (r) or Spearman (rho) correlation coefficients. Furthermore, a two-way repeated-measures mixed design ANOVA (group × intensity) was used to compare the change in fat oxidation rates with exercise intensity .Data are expressed as the mean ± SD for all variables. The sample size of 11 participants per group was calculated based on previously published data . Before p = 0.40 and p = 0.36, respectively). In comparison with the active group, cyclists had a lower body mass index (BMI) (p = 0.02) and FM (in % and in kg) (p = 0.02 for both), while their p < 0.001), confirming that the aerobic fitness level was different between the two experimental groups. Although no difference was found in the RCP and VT expressed as % p = 0.40 and p = 0.09, respectively), the absolute values of the VT and RCP were higher in cyclists (p < 0.001).The participants’ physical characteristics and their aerobic fitness parameters are listed in –1⋅min–1) was higher in the cyclist group (p = 0.03) than that in the active group for the exercise intensities between 20 and 65% p ≤ 0.046; max in mL⋅kg–1⋅min–1 were higher in cyclists than in active individuals (p = 0.01 for both). However, no difference was found when Fatmax was expressed as %p = 0.77). The dilatation was greater in the active group than in the cyclist group (p = 0.003), while no difference in symmetry or translation was found between the two groups .Whole-body fat oxidation kinetics, expressed in relative and absolute values, are shown in r = 0.65; p = 0.001), MAP , and VT in mL⋅kg–1⋅min–1  and in %r = 0.68; p = 0.001) and was correlated with RCP only when expressed in mL⋅kg–1⋅min–1 . Similarly, Fatmax (mL⋅kg–1⋅min–1) was also positively correlated with r = 0.73; p < 0.001) and VT in mL⋅kg–1⋅min–1  and in %r = 0.59; p = 0.004) and was correlated with RCP only when expressed in mL⋅kg–1⋅min–1 . In contrast, the relative value of Fatmax was not correlated with any aerobic fitness parameters.Maximal fat oxidation was positively correlated with BP in cyclists compared to that in active individuals. In fact, a1 was lower in the cyclist group than in the active group (p = 0.02). The [HHb]BP occurred at a higher absolute intensity (in mL⋅kg–1⋅min–1) but not at a higher relative intensity . The [HHb]BP expressed in mL⋅kg–1⋅min–1 was strongly and positively correlated with r = 0.88; p < 0.001), MAP , VT , and RCP .The %Δ[HHb] response as a function of relative and absolute exercise intensity is shown in a1 (p = 0.06) and was positively correlated with [HHb]BP (p = 0.01). Fatmax (mL⋅kg–1⋅min–1) tended to be negatively correlated with a1 (p = 0.06) and was positively correlated with a3 (p = 0.03), [HHb]BP (p < 0.001). When expressed in relative values (%max was only positively correlated with a3 (p = 0.01). Finally, the symmetry was positively correlated with a3 (p = 0.01). A schematic view of the correlations among aerobic fitness, whole-body fat oxidation and muscle deoxygenation kinetics is presented in The correlations between whole-body fat oxidation and muscle deoxygenation parameters are listed in max and (2) shallower muscular deoxygenation kinetics  with a right-shifted [HHb]BP compared to active participants. MFO and Fatmax were both positively correlated with the [HHb]BP and tended to be negatively correlated with a1  significant upward fat oxidation kinetics with greater MFO and absolute Fat with a1 .max (in mL⋅kg–1⋅min–1) was also higher in cyclists than in the active group (+29%), whereas no difference was found for Fatmax when expressed as relative intensity   , 2012, w1⋅min–1) , 2015. A1⋅min–1) , the dil designs . However2 delivery and consumption, reflected by a slower slope (a1) of %Δ[HHb] kinetics (-20%) and a [HHb]BP occurring at a significantly higher absolute intensity (+42%) compared with the active group . In other words, whole-body fat oxidation kinetics presenting a rightward asymmetry; thus, a right-shifted Fatmax may be associated with a steeper slope of the second part of the %Δ[HHb] kinetics. This may reflect a prolonged reliance on aerobic metabolism and thus a prolonged utilization of fat as an energetic substrate, even at high intensity when glycolytic metabolism provides more energy during exercise compared with lower intensities. In fact, our a3 values are different from zero, attesting that, at the end of the submaximal incremental test, the upper limits of the fractional O2 extraction were not yet reached  and Fatmax shifted to higher absolute exercise intensities. This corroborates that, compared to active individuals, endurance-trained cyclists may maintain aerobic ATP production until higher exercise intensities and may have a delayed reliance on glycolytic metabolism because of a longer maintained balance between O2 delivery and consumption, supporting higher fat oxidation rates.Our findings confirmed the influence of aerobic fitness status on both whole-body fat oxidation  and muscThe datasets generated for this study are available on request to the corresponding author.The studies involving human participants were reviewed and approved by Commission cantonale d’Éthique de la Recherche sur l’être humain Vaud (CER-VD)—Swissethics. The patients/participants provided their written informed consent to participate in this study.AZ, XC, and DM conceived and designed the research study. AZ, SL, CT, NB, XC, and DM performed the experiments. AZ, SL, LV, CT, XC, and DM analyzed the data. AZ, BG, BK, XC, and DM interpreted the results of the experiments. AZ, XC, and DM prepared figures and drafted the manuscript. All authors approved the final version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


According to the electrochemical measurements, the material presents an ultrahigh capacitance of 374 F g−1 (the current density is 1 A g−1). Furthermore, the material demonstrates excellent rate performance (105 F g−1 at a higher density of 20 A g−1) and ideal cycling stability (87.3% capacity retention after 5000 times charge–discharge at 2 A g−1). When used for a symmetrical supercapacitor device, the material also shows a relatively high capacity of 330 F g−1 at 1 A g−1 (a two-electrode system). All measurements suggest the material is an effective and noteworthy material for the electrodes of supercapacitors.Carbon materials have been widely used as electrode materials for supercapacitors, while the current carbon precursors are mainly derived from fossil fuels. Biomass-derived carbon materials have become new and effective materials for electrodes of supercapacitors due to their sustainability, low pollution potential, and abundant reserves. Herein, we present a new biomass carbon material derived from water hyacinth by a novel activation method (combination of KOH and HNO The exploration of efficient and clean devices for energy storage has become an essential challenge for human beings. Supercapacitors have been considered to be promising candidates with advantages of long cycle life, high power density, wide working temperature range, high charge–discharge efficiency, and stability ,2,3,4,5.Green biomass-derived carbon materials have raised hopes because of their abundant reserves, low pollution, sustainability, and applicability. To date, many kinds of biomass sources including animals and plants have been explored as precursors for carbon-based electrode materials ,25,26,273 activation) is reported, which shows a superb electrochemical capacitance performance for supercapacitors. Generally, the HNO3 activation process is first undertaken after a direct carbonization treatment (the sample prepared by direct carbonization denoted as water hyacinth-derived carbon: WHC), some pores are generated as well as the SSA value is increased, and the obtained material by this activation is denoted as activated water hyacinth-derived carbon (AWHC). Subsequently, the KOH activation process was undertaken, in which vast micropores and mesopores could be generated. The obtained final sample which is denoted as double activated water hyacinth-derived carbon (AAWHC) possesses an ultrahigh SSA of 1622.6 m2 g−1 and vast mesopores and micropores. AAWHC shows an ultrahigh specific capacitance of 374 F g−1  at 1 A g−1 and a long cycle life of 87.3% retention (5000 times charge-discharge) at a current density of 2 A g−1.Herein, a new biomass carbon derived from water hyacinth by an ordered activation method  (1 g:20 mL) were stirred at normal temperature for a whole day. Next the achieved mixtures were washed several times with deionized water, then heat treated in a tubular furnace at 800 °C for 2 h. After cooling to normal temperature, samples were treated by washing and drying processes to obtain AWHC, the above corresponds to step one of the activation process. The following is step two of the activation process. Mixtures of AWHC and KOH  (a mass ratio of 1:3) were fully ground and then treated in a tubular furnace at 800 °C for 2 h. After cooling to room temperature, 1 M HCl  aqueous solution was mixed with the sample with stirring for 4 h to wash away the redundant KOH and then rinsed repeatedly with deionized water until the pH changed to 7.0, before being dried to achieve the final sample AAWHC . An amount of 100 g water hyacinth obtained from a pond was rinsed more than once with absolute ethyl alcohol and deionized water, and dried in an oven at 70 °C for 24 h to obtain the dried water hyacinth leaves. Subsequently, the leaves were carbonized directly at 800 °C for 12 h to acquire the WHC. Then the mixtures of WHC and 98% HNO−1). The element compounds were measured using X-ray photoelectron spectroscopy  with Al Ka excitation (1486.6 eV). Raman patterns were tested using a laser Raman microscope  (wave-length of argon ion laser: 633 nm). The pore properties were measured with a Barrett–Joyner–Halenda (BJH) method and SSA was detected with a Brunauer–Emmett–Teller (BET) method. Scanning electron microscope  and transmission electron microscope  were conducted to analyze the microcosmic morphology and structures. Differential scanning Calorimetry (DSC) and Thermogravimetry (TG) tests were conducted by a thermal analyzer  at a temperature of 0–1000 °C  and carbon black in mortar (mass ratio: 8:1:1), then pure ethanol was mixed, and the mixture ground adequately to achieve a slurry before smearing immediately on a pure nickel foam. The nickel foam was washed by HCl, deionized water, and absolute ethyl alcohol several times with ultrasound to ensure purity. The foams were dried in an oven at 120 °C for a whole day and treated with a tableting process to obtain the working electrodes.−1. CV tests were conducted with a potential range of −0.8 to 0.2 V vs. Ag/AgCl electrode at different scan rates of 5, 10, 20, 50, 100 mV s−1. The EIS tests were conducted with a frequency range of 0.01Hz to 100 KHZ (amplitude: 5 mV). Moreover, a simple supercapacitor device was constructed by utilizing two identical AAWHC electrodes and measured with a two-electrode system in 6M KOH solution. Besides, cycle measurements, the values of the charge and discharge time were calculated (5000 cycles at 20 A g−1). In a three-electrode system, capacitance −1) is the scan rate, Measurement conditions: For the measurements of capacitance behaviors of WHC, AWHC, and AAWHC, the electrochemical workstation CHI760 was utilized for galvanostatic charge–discharge (GCD), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) measurements. Specifically, an Ag/AgCl electrode was prepared as reference electrode and a platinum film electrode as counter electrode, the electrolyte was 6 M KOH aqueous solution. In a three-electrode system, the GCD measurements were conducted with a potential range of −0.8 to 0.2 V at current densities of 1, 2, 5, 10, 20 A g−1) is the capacity value, −1 ) is the specific power density, and −1 ) is the specific energy density.For these assembled devices, capacity value 3 activation precedes KOH activation). This scheme was also used for presenting the variations in structure  and porosity (increase of pores). The HNO3 activation process promotes some complex reactions which can increase the polarity, pores and SSA, indicating sufficient ion transmission channels and electrochemical active sites [The preparation processes are shown in ve sites ,45. The ve sites ,47,48. T3 activation and KOH activation. As shown in 3 activation; no vast pores are generated to significantly change the structure morphology. As shown in SEM and TEM were used to explore the morphology and microstructure of samples. As shown in 2 adsorption/desorption system. As shown in 2 g−1 while the value of AWHC is 740 m2 g−1 and WHC is 350 m2 g−1. AAWHC possesses a higher mesopore content (0.3962 mL g−1) than that of AWHC (0.1127 mL g−1) and WHC (0.0856 mL g−1), which means the pore size distribution of AAWHC is more reasonable. When used as electrode materials, sufficient micropores and mesopores as well as high SSA can provide sufficient electrolyte ion transmission channels and active sites for electrochemical reactions, further accelerating electrode efficiency [The SSA and pore properties of all samples were measured by the Nficiency ,53,54.−1 and G (graphite) peak at ~1588 cm−1 [D/IG of AAWHC (1.01) is higher than that of WHC (0.88) and AWHC (0.93), indicating that AAWHC samples contain more edges and defect sites, which are beneficial for electrochemical reactions [The crystalline structures of the samples were confirmed by analyzing the XRD patterns. According to 588 cm−1 . The inteactions ,61. −1 (−1) and the values for AWHC and WHC are 114 F g−1 and 47 F g−1, respectively. However, the pure nickel foam possesses a basic capacitance, so the pure nickel foam underwent the additional GCD test—the measured capacitance value suggests this has no influence on the accuracy of the calculation results . All the results demonstrate that AAWHC offers excellent energy density and capacity behaviors, probably due to the excellent surface chemical properties and structural characteristics of AAWHC. Two identical AAWHC electrodes were constructed in a simple symmetrical supercapacitor device (electrolyte: 6.0 M KOH solution). According to the CV curves of the AAWHC-based device a, which 20 A g−1 c, and thng to −1 d, the ca3 activation and KOH activation. The as-prepared materials display a loose bulk structure with abundant channels and some wrinkles. Acting as electrode material for a supercapacitor, AAWHC shows a superb specific capacity of 374 F g−1 at a 1 A g−1 and a promising rate performance. Moreover, AAWHC shows a high specific energy density of 11.46 Wh kg−1 and an excellent cycle life (87.3% capacity retention after 5000 times charge and discharge) in a symmetric supercapacitor device. It is certain that with the superiority displayed in this paper, such a mesopore-rich carbon material which possesses advantages of low-cost and sustainability will have promising potential for energy storage applications.To summarize, a new mesopore-rich nanoporous biomass carbon AAWHC derived from water hyacinth was fabricated by an innovative preparation method which combined HNO


Sophora japonica ‘JinhuaiJ2’ (Papilionaceae), known as Chinese Scholar Tree, is an important traditional herb with a long history of cultivation in China. It’s well known for its valuable medicinal values due to its flower buds contains abundant rutin. Here, we reported and characterized its complete chloroplast genome based on Illumina paired-end sequencing data. The complete plastid genome was 158,690 bp, which contained inverted repeats (IR) of 25,339 bp separated by a large single-copy (LSC) and a small single copy (SSC) of 88,978 bp and 19,034 bp, respectively. The cpDNA contains 129 genes, comprising 83 protein-coding genes, 38 tRNA genes and 8 rRNA genes. The overall GC content of the plastome is 36.1%. The phylogenetic analysis of 18 selected chloroplast genomes demonstrated that S. japonica ‘JinhuaiJ2’ was close to the species Tapiscia sinensis. Sophora japonica ‘JinhuaiJ2’ (Papilionaceae), which belongs to the Leguminosae family, is an important traditional Chinese herb with a long history of cultivation  was deposited in the Guangxi Key Laboratory of Plant Conservation and Restoration Ecology in Karst Terrain, Guangxi Institute of Botany, Guangxi Zhuang Autonomous Region and Chinese Academy of Sciences, Guilin, China. The experiment procedure is as reported in Zhang et al. (http://www.ncbi.nlm.nih.gov/.).In this study, g et al. . Around g et al.  and direg et al.  and SequS. japonica ‘JinhuaiJ2’ is a typical quadripartite structure with a length of 158,690 bp, which contained inverted repeats (IR) of 25,339 bp separated by a large single-copy (LSC) and a small single copy (SSC) of 88,978 bp and 19,034 bp, respectively. The cpDNA contains 129 genes, comprising 83 protein-coding genes, 38 tRNA genes and 8 rRNA genes. Among the annotated genes, 16 of them contain one intron , and two genes (clpP and ycf3) contain two introns. The overall GC content of the plastome is 36.1%.The chloroplast genome of S. japonica ‘JinhuaiJ2’, phylogenetic analysis was conducted. A neighbor joining (NJ) tree with 1000 bootstrap replicates was inferred using MEGA version 7 (Kumar et al. S. japonica ‘JinhuaiJ2’ was close to the species Tapiscia sinensis (To identify the phylogenetic position of sinensis . Our fin


In epithelial ovarian cancer (EOC), the strongest prognostic factor is the completeness of surgery. Intraoperative molecular imaging that targets cell-surface proteins on tumor cells may guide surgeons to detect metastases otherwise not visible to the naked eye. Previously, we identified 29% more metastatic lesions during cytoreductive surgery using OTL-38, a fluorescent tracer targeting folate receptor-α (FRα). Unfortunately, eleven out of thirteen fluorescent lymph nodes were tumor negative. The current study evaluates the suitability of five biomarkers  as alternative targets for molecular imaging of EOC metastases and included FRα as a reference. Immunohistochemistry was performed on paraffin-embedded tissue sections of primary ovarian tumors, omental, peritoneal and lymph node metastases from 84 EOC patients. Tumor-negative tissue specimens from these patients were included as controls. EGFR, VEGF-A and L1CAM were highly expressed in tumor-negative tissue, whereas αvβ6 showed heterogeneous expression in metastases. The expression of EpCAM was most comparable to FRα in metastatic lesions and completely absent in the lymph nodes that were false-positively illuminated with OTL-38 in our previous study. Hence, EpCAM seems to be a promising novel target for intraoperative imaging and may contribute to a more reliable detection of true metastatic EOC lesions. Epithelial ovarian cancer (EOC) is the second most common cause of death among all gynecological malignancies, leading to 125,000 deaths per year worldwide .The standard treatment for patients with apparent early-stage EOC is a staging procedure consisting of the removal of both ovaries, fallopian tubes, uterus, omentum and lymph nodes and careful the examination of surrounding tissues for the presence of metastatic spread. If no suspicious lesions are detected, biopsies are taken from normal-appearing tissue in predefined areas to determine the extent of disease and the possible need for additional chemotherapy . For patA leading example of tumor-targeted molecular imaging is near-infrared fluorescence (NIRF) imaging. NIRF imaging can be used intraoperatively to accurately discriminate malignant from benign tissue in real time using fluorescent probes. Ideally, an NIRF probe consists of targeting moieties such as antibodies, peptides or ligands which bind with high affinity to proteins or receptors that are overexpressed on the cell surface of tumor cells but are absent on adjacent normal cells. These targeting moieties are conjugated to a fluorophore which emits light in the near-infrared range λ = 700–900 nm) [00 nm 9,. The tumTo improve the outcome of cytoreductive surgery by means of tumor-targeted imaging, proteins that are exclusively overexpressed on the cell surface of EOC cells should be identified . Folate This study examines the expression of biomarkers to determine their value as novel targets for the NIRF imaging of metastatic EOC lesions with similar accuracy to FRα, but with improved discrimination between tumor-positive and tumor-negative tissue. Recent studies have shown great improvements in understanding the molecular landscape of epithelial ovarian cancer . We focun = 485 samples) from the same patients, were included. Sections from these samples were evaluated by immunohistochemistry to assess expression patterns for EGFR, VEGF-A, L1CAM, EpCAM, integrin αvβ6 and FRα. Tissue samples were derived from 42 patients who underwent either primary cytoreductive surgery (n = 297 samples) or 42 patients who underwent interval cytoreductive surgery (n = 188 samples). Subgroups were divided based on histological subtype according to the classification model by Shih and Kurman [n = 73) and “other histological subtypes” (n = 11), representing low grade serous carcinoma (n = 1), endometrioid adenocarcinoma (n = 4), clear cell carcinoma (n = 2) and mucinous adenocarcinoma (n = 4). Patient and tumor characteristics are shown in In total, 84 primary tumors, 70 omental, 86 peritoneal, 11 lymph node metastases and 12 tumor samples from diagnostic biopsies (ascites) before NACT derived from 84 patients with EOC, as well as 222 tumor-negative samples .The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and area under the curve (AUC) were calculated for EpCAM, αvβ6 and FRα based on either “overexpression” or “no overexpression” to determine the potential of correctly identifying tumor-positive and tumor-negative lymph nodes in primary debulking samples . αvβ6 shTo determine whether chemotherapy can affect biomarker expression, tissue samples from both primary and interval cytoreductive surgery were included. When comparing both TIS and intensity scores, there was no statistically significant difference in αvβ6 and FRα expression between primary and interval debulking. However, there was a significant difference in EpCAM expression between both groups in metastases, but not in the primary tumors.n = 12) before start of NACT from patients in the interval cytoreductive surgery group were immunohistochemically stained for EpCAM, αvβ6 and FRα. Expression patterns before NACT were compared to the remaining vital tumor cells sampled during interval cytoreductive surgery. Although a small decrease was seen in staining intensity for all targets after NACT in the intensity scores bar graphs . In line with this, the homogenous expression of a biomarker will result in a better demarcation of metastatic deposits than heterogenous expression patterns, making EpCAM and FRα more suitable targets for intraoperative imaging than αvβ6.Although EpCAM and FRα meet the criteria of a suitable molecular target, high expression was observed in epithelial cells in tumor-negative fallopian tubes and endometrial cells of the uterus. Yet, this will not interfere with surgical performance using intraoperative imaging, as these structures are routinely removed during both surgical staging and cytoreductive surgery. This would only pose a challenge for the application of NIRF-guided surgery in women with early-stage EOC who wish to preserve their fertility. In these patients, the uterus and contralateral ovary and fallopian tube will be left in situ in case of normal appearance . As a reSimilarly, EpCAM expression was observed in epithelial cells lining the intestinal lumen. Since an NIRF probe has a tissue penetration depth of up to 1 cm, these cells may be detected by a probe targeting EpCAM and result in background fluorescence. On the contrary, it is possible that EOC metastases located on the serosal surface of the small bowel can still be adequately discriminated. After all, it has been shown that there are four times as many EpCAM molecules expressed on a malignant cell compared to a normal cell, and while this may not be observed by immunohistochemistry, a difference in fluorescence intensity could be perceived . Hence, Strikingly, no expression of EpCAM, αvβ6 or FRα was observed in the lymph nodes that were previously false-positively illuminated with OTL-38. It has been suggested that the false positive signal detected in the lymph nodes was likely due to the fact that OTL-38 not only binds to FRα, but also FRβ, which is expressed by tumor-associated macrophages (TAMs) present in the lymph nodes. The presence of FRβ in the previous false positive lymph nodes was confirmed via immunostaining . Since, Increases in EpCAM expression have been correlated to cancer development and progression and increased proliferation ,26,27, wwww.clinicaltrials.gov, NCT03399344). Another promising technique to preoperatively assess metastatic tumor load in patients with EOC might be a pre-operative tumor-targeted positron emission tomography (PET) scan using tumor-specific radioactive tracers. To use EpCAM as a target for tumor-specific PET imaging, antibodies or peptides against EpCAM can be conjugated to a radioactive isotope. A combination of tumor-specific PET and NIRF imaging could further improve the detection of metastatic lesions.Beside the intraoperative use, tumor-targeted molecular imaging may also be used pre-operatively to determine tumor load and surgical resectability. The pre-operative estimation of surgical resectability in EOC is currently based on computed tomography (CT) scans. However, a CT scan may underestimate the extent of disease. As a result, a significant proportion of patients with advanced stage disease will undergo a futile laparotomy ,29,30. Bn = 84), their corresponding omental, peritoneal and lymph node metastases (n = 167) and tumor tissue from diagnostic biopsies before NACT, if applicable (n = 12). Tumor-negative tissue specimens in the primary debulking group originated from patients who underwent a staging procedure because of suspected early-stage EOC. Tumor-negative tissue specimens in the interval debulking group originated from biopsies taken during interval debulking procedures and were therefore subject to NACT. To determine expression in tumor-negative tissue from surrounding structures, samples from the ovaries, fallopian tubes, uterus, omentum, pouch of Douglas, bladder peritoneum, pelvic wall, paracolic gutter, diaphragm, intestine/appendix and lymph nodes were included from all patients (n = 222). Subjects who were alive at the time of the study gave their informed consent for the use of their tissue. This study was conducted in accordance with the Declaration of Helsinki and approved by the Ethics Committee (protocol number B17.025).Medical records and tissue specimens were retrospectively reviewed from patients diagnosed with advanced stage EOC (FIGO IIb-IV ) who undn = 5) and tumor-negative peritoneal (n = 3), omental (n = 3) and lymph node (n = 3) tissue.Because previous research has indicated heterogenous expression levels of EGFR, VEGF-A and L1CAM in EOC ,20,32, an = 11) were immunohistochemically stained for the most promising targets.Additionally, considering the aforementioned false-positivity in lymph nodes reported in a translational study with an FRα targeting probe (OTL-38) by our group , these fWhole formalin-fixed paraffin-embedded (FFPE) tissue blocks were cut into sections of 4 µm thickness. After the deparaffinization of these sections in xylene and rehydration in a stepwise series of alcohol solutions, endogenous peroxidase activity was blocked with 0.3% hydrogen peroxide for 20 min. Deparaffined slides were placed in a 37 °C water bath and incubated with 0.125% trypsin for 30 min to unmask EpCAM epitopes, and with 0.4% pepsin and 1N HCl for 20 min to unmask αvβ6 and EGFR epitopes. VEGF-A epitopes were unmasked by heat induction at 95 °C using citrate buffer  in a PT-Link module . Antigen retrieval for L1CAM was performed by a 30 min incubation in Tris-EDTA  pre-heated to 90–100 °C. The slides were then incubated overnight in a humidified chamber at room temperature with previously determined dilutions of primary monoclonal antibodies against EpCAM (0.25 µg/mL), αvβ6 (0.5 µg/mL), EGFR (2.86 µg/mL), VEGF-A (0.25 µg/mL) and L1CAM (1 µg/mL). The slides were then washed in PBS and incubated for 30 min at room temperature with a horseradish peroxidase (HRP)-labelled secondary antibody . Staining for FRα was performed using an immunohistochemical assay kit according to the manufacturer’s instructions . After being rinsed in PBS, the immunoreactions were visualized using DAB substrate buffer (Dako) for 10 min and counterstained using Mayer’s hematoxylin for 15–30 s. After dehydration at 37 °C, the stained slides were mounted with pertex . All antibodies and reagents used for immunohistochemical staining can be found in All immunohistochemically stained slides were scanned using the Ultra-Fast Scanner 1.6 RA . Staining in malignant tissues was evaluated based on membranous expression. Positive expression was scored independently by two blinded observers (L.M. and M.D.), of which a final agreement score was determined. Samples with no agreement were resolved by consensus. The score comprised the total amount of positively stained tumor cells as a percentage of total tumor tissue (proportion score (PS): 0 ≤ 9%, 1 = 10–25%, 2 = 26–50%, 3 = 51–75%, 4 ≥ 76%), and the staining intensity (intensity score (IS): 0 = none, 1 = weak, 2 = moderate, 3 = strong). Examples of staining intensities can be seen in p < 0.05. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and area under the curve (AUC) were calculated for all tumor-negative and tumor-positive lymph nodes for EpCAM, αvβ6 and FRα. Calculations were based on the TIS scores and were classified as “overexpression” (TIS 6–12) and “no overexpression” (TIS 1–4). Tumor-negative lymph nodes with no observed expression were classified as “no overexpression”. Sensitivity was calculated by dividing the lymph nodes with true overexpression by the total number of tumor-positive lymph nodes. Specificity was calculated by dividing the lymph nodes with true no overexpression by the total number of tumor-negative lymph nodes. PPV was calculated by dividing the lymph nodes with true overexpression by the total number of lymph nodes with overexpression, and NPV was calculated by dividing the lymph nodes with true no overexpression by the total number of lymph nodes with no overexpression. The AUC was calculated using SPSS. Further graphical representations of staining intensities were created using GraphPad Prism 7 . Image representations of immunohistochemical stainings were created using Adobe Illustrator CC 2018 .All statistical analyses were performed using SPSS software version 23 . Agreement scores were calculated using Cohen’s kappa. A one-way ANOVA with a post-hoc Bonferroni correction was applied to calculate mean percentage staining and the difference in staining between tissues. Results were considered statistically significant if In conclusion, our study showed that EpCAM is a potential alternative target for FRα for tumor-specific intra-operative molecular imaging for EOC. Since a high and homogenous expression was detected in primary tumors and metastases, and no false positivity in lymph nodes was observed, EpCAM may contribute to a more reliable detection of metastatic EOC lesions. Further research is needed to determine whether guidance by tumor-targeted molecular imaging will more often lead to the completeness of surgery.


Diabetes mellitus is a chronic burden, with a prevalence that is increasing worldwide. Telemetric interventions have attracted great interest and may provide effective new therapeutic approaches for improving type 2 diabetes mellitus (T2DM) care.1c (HbA1c) specifically and T2DM management generally in a systematic meta-review.The objective of this study was to analyze the clinical effectiveness of telemetric interventions on glycated hemoglobin A1c, blood pressure, fasting blood glucose, BMI, diabetes-related and health-related quality of life, cost-effectiveness, time savings, and the clinical effectiveness of telemetric interventions were analyzed. In total, 73 randomized controlled trials (RCTs), 10 systematic reviews/meta-analyses, 9 qualitative studies, 2 cohort studies, 2 nonrandomized controlled studies, 2 observational studies, and 1 noncontrolled intervention study were analyzed.A systematic literature search was performed in PubMed, CINAHL, Cochrane Library, Web of Science Core Collection, and EMBASE databases from January 2008 to April 2020. Studies that addressed HbA1c, suitable RCTs were pooled and the average was determined. An HbA1c decrease of –1.15% (95% CI –1.84% to –0.45%), yielding an HbA1c value of 6.95% (SD 0.495), was shown in studies using 6-month “real-time video” interventions.Overall, 1647 citations were identified. After careful screening, 99 studies  were selected by two independent reviewers for inclusion in the review. Telemetric interventions were categorized according to communication channels to health care providers: (1) “real-time video” interventions, (2) “real-time audio” interventions, (3) “asynchronous” interventions, and (4) “combined” interventions. To analyze changes in HbA1c values in both the short term and the long term and contribute to the effective management of T2DM. More studies need to be done in greater detail.Telemetric interventions clearly improve HbA Diabetes mellitus is a chronic burden, with a prevalence that is increasing worldwide . In 2019T2DM is closely associated with diabetic microvascular complications—such as nephropathy, retinopathy, and neuropathy —and macrTherefore, optimal glycemic management is crucial . Recent 1c (HbA1c), blood pressure (BP), fasting blood glucose (FBG), BMI, diabetes-related quality of life (DRQoL), and health-related quality of life (HRQoL), as well as the cost-effectiveness, time savings, and clinical effectiveness of telemetric interventions in general. HbA1c is one of the major clinical parameters in T2DM and therefore our main focus.In this systematic meta-review , we focuTo our knowledge, this study is the first and only systematic meta-review of telemetric interventions in T2DM management with respect to the following special features: we developed and applied a unique classification system for analyzing telemetric interventions and provide detailed insights by including several study designs and a wide range of clinical outcomes.A systematic search was conducted targeting the period between January 2008 and April 2020. No protocol has been published. Keywords  were selected from the MEDLINE Medical Subject Headings and EMBASE Subject Headings databases and searched in titles/abstracts . In genePublications addressing telemetric interventions targeting T2DM management were included.Telemetry was defined as “a mode of delivering healthcare services through the use of telecommunications technologies, including but not limited to asynchronous and synchronous technology, and remote patient monitoring technology, by a healthcare practitioner to a patient or a practitioner at a different physical location than the healthcare practitioner” . We inclStudies were screened and selected by two independent reviewers. Disagreements were resolved by a consensus-based discussion. We selected studies that met the following inclusion criteria: (1) peer-reviewed articles and studies; (2) written in English or German; (3) study design was an SR, MA, CT, or RCT; and (4) included interventions that involved direct interaction between patients and health care professionals through feedback and data transmission. We also considered quantitative and qualitative studies.Smartphone/mobile app–based interventions were excluded and analyzed separately in another publication. We also rejected publications that observed mixed populations , provided pooled data with other digital applications, addressed prevention or diagnosis, or focused on the presentation of technologies. Year of publication, location of the study, duration of the intervention, study design, sample sizes, intervention and control groups used, frequency of contact, feedback methods, outcomes, effects, statistical significance, and conclusions were extracted from each publication (step 7).For analysis, the interventions were classified a priori (step 8) based on the technologies used, study design, and outcomes .“Real-time video” interventions (12 intervention studies): synchronous, face-to-face communication by videoconferencing and video consulting.“Real-time audio” interventions (17 intervention studies): synchronous communication by telephone calls (telephone coaching and counselling).“Asynchronous” interventions (28 intervention studies): asynchronous communication by email, SMS text messaging, internet/web-based platforms, server, home gateway, and post.“Combined” interventions (33 intervention studies): interventions involving real-time  and asynchronous communication, with a subgroup of “video clips” .1c concentrations, differed. To determine the change in HbA1c, we pooled appropriate RCTs and calculated the differences in means and 95% CIs for the intervention and control groups at the study end points. RCTs in which the changes from baseline to the end of the study were reported as a percentage were included. Studies in which the control group received telemetric support were excluded. Mean deviations and SDs were extracted unchanged.We conducted a small subgroup MA to assess whether the impact of the four intervention types, as well as the short- and long-term effects on the management of HbA1c values based on the RCTs and the mean differences (MDs) from our subgroup MA.In addition, the publication bias was assessed visually as a funnel plot using HbAWe also pooled the number of patients, specifically the number of unique patients as well as the number of patient cases related to the outcomes. In the former scenario, each patient occurred only once, addressing the number of individual patients (without SRs and MAs), and in the latter scenario, with a focus on specific outcomes, patient cases were analyzed based on the respective outcomes and thus may have been included several times (including SRs and MAs).Our search strategy identified 1647 citations. After removing duplicates, 1116 studies were screened and 875 ineligible papers excluded. After assessing 241 studies with full text, 72 inappropriate studies were rejected. As an interim result, 189 studies were identified, of which 23 focused on type 1 diabetes mellitus, 99 focused on T2DM, 11 focused on gestational diabetes, and 51 focused on mixed populations. In this systematic meta-review, we included 99 suitable T2DM publications, analyzing 15,939 patients and 82,439 patient cases. A list of the included studies is provided in Baseline characteristics of the studies are summarized in In total, 12 “real-time video,” 17 “real-time audio,” 28 “asynchronous,” 33 “combined,” and 3 “video clip” interventions were classified. One study matched the classification criteria for two categories . A detai1c values indicated a mild form of asymmetry  by implementing telemetric interventions [1c values compared with the usual care .I²=16.7%) compared with usual care.According to Kim et al , telemonDue to the heterogeneous data situation, Zhai et al  could no1c values. More specifically, five RCTs  indicated significant positive effects  [Overall, 89% (8/9) of the studies reported a clear reduction in HbA , wherea [P<.015  providedP>.05 [P=.76; diastolic BP 72.7 mmHg, SD 2.1 mmHg, versus 70.2 mmHg, SD 2.2 mmHg, P=.64).In general, most RCTs observed no essential changes in systolic and diastolic BP measurements (intergroup P>.05  and signP>.05 ). HoweveP>.05 , althougP=.023).Rasmussen et al  showed aP=.73).Hansen et al  found noHansen et al  noted thGordon et al  revealedCarlisle and Warren  suggeste1c levels with audio interventions in real time. Odnoletkova et al [P=.003 [P=.04 [1c levels compared with control groups (P>.05). Notably, some control groups [1c concentrations (P<.05).In summary, all studies showed precise improvements in HbAva et al  and Walkva et al  reported [P=.003 ; and int3 [P=.04 ). Saraya3 [P=.04 , Trief e3 [P=.04 , Maslakp3 [P=.04 , Blackbe3 [P=.04 , Benson 3 [P=.04 , and Vas3 [P=.04  displayel groups -29,31 rel groups  found th1c decreased at a rate of 0.32% every 3 months for the online arm, 0.36% for the telephone arm, and 0.41% for the web training arm .The RCT of McMahon et al  was clasP=.02), but not in a long-term way, while Maslakpak et al [P=.766).The “real-time audio” intervention studies showed obvious improvements in FBG. In the study by Varney et al , FBG levak et al  outlinedP=.021). Vasconcelos et al [P=.171) and diastolic BP . In addition, Varney et al [In general, all “real-time audio” intervention studies reported clear improvements in BP. Trief et al  showed aos et al  reportedey et al  indicateP=.004).According to Odnoletkova et al , the difP=.003 [P=.021 [P=.764).In general, all trials noted slight improvements in BMI. Odnoletkova et al  and TrieP=.003  and P=.0 [P=.021 ), wherea [P=.021  indicate“Real-time audio” interventions appear to be moderate in terms of cost-effectiveness . Schechter et al  conclude1c in the intervention groups compared with the control groups (P<.05) [P<.05) [P>.05) [P=.004 [P<.001, versus internet blood glucose monitoring system, P<.05 [P<.01 [P>.05).Overall, the majority of the studies  reported apparent improvements. Eleven RCTs reported significant improvements in HbA (P<.05) -46, wher [P<.05) -49. In a [P>.05) -54. Rama [P>.05) , Tildesl [P>.05) , and Cho [P>.05)  mentione [P=.004 ; real-tim, P<.05 ; and P<.5 [P<.01 ), but thP=.015 [P=.005 [The studies showed significant improvements in FBG levels in the intervention groups compared with control groups .P=.017) and diastolic BP (P=.006) in the intervention group compared with the control group. Wakefield et al [P=.01). Fang and Deng [P=.069) and diastolic BP (P=.693) in the treatment group, but they were not significant.In summary, the publications reported apparent improvements in BP. Wild et al  found sild et al  also fouand Deng  found imP=.000) and BMI .In general, the studies revealed clear beneficial effects of “asynchronous” interventions on both body weight and BMI. For example, Luley et al  showed lNo clinically important improvements in HRQoL were seen according to Dario et al .A weight-loss telemonitoring intervention from Luley et al 2011  showed a1c levels greater than 6.5% (P<.05).Cho et al  showed a1c (P<.05). Three of these studies were RCTs that achieved significant improvements within their intervention groups (P<.001 [P=.27 [P>.05).In general, most publications  reported clear significant improvements in HbA  but notP<.001 [P=.027 .P<.01 [P=.035, and diastolic BP, P=.013 [P<.05) but no significant differences between the groups (P>.05) [P<.01 [P<.001 [Approximately 85% (11/13) of the “combined” intervention studies reported beneficial effects on BP -61,63-70, P=.013 ). Howeve (P>.05) ,66-69,71P<.01).Most “combination” intervention studies  found clear improvements in body weight ,69,70,72P=.047 [P<.01 [P<.001 [P=.036 [P<.01 [P<.05 [P<.01) and Kempf et al [P<.01).The majority of publications  showed an apparent reduction of BMI. Significant improvements were outlined by 6 studies . For ex1 [P<.05  . Jha et al [P=.015 and P<.001, respectively).All studies found clear improvements in DRQoL and HRQoL. Kempf et al  and Nicoha et al  and Dienha et al  . AccordP<.001 , the extHsu et al  reported1c compared with control subjects .P=.306, and diastolic BP, P=.374) but no effects on body weight (P=.232).Tang et al  detected1c values between the intervention and control groups at the study end points were investigated. Patients’ changes in HbA1c from baseline to the end of the study of 41 RCTs are presented in Short- and long-term effects based on the comparison of HbAA small MA showed that, compared with the control group, 6-month interventions (n=2) were associated with a greater effect size  than 12-month interventions (n=2) .The subgroup analysis revealed an effect size, compared to usual care, of MD=–0.37% (95% CI –0.79 to 0.05) for 6-month interventions (n=3) compared with –0.5% in the 3-month intervention  and –0.0The greatest effect was seen in 12-month interventions (n=2) , followed by 6-month interventions (n=8) , and 3-month interventions (n=3) .The 3-month interventions (n=5) had the greatest effect , whereas 6-month interventions (n=7) had a slightly smaller effect . In comparison, the effect of 12-month interventions (n=4) was even smaller . The subgroup “video clip” interventions (n=2) showed a reduction of MD=–0.23 (95% CI –0.23 to –0.23).1c levels [1c levels. Telemetry has the advantage of helping people who are restricted due to geographic location or a lack of resources [Telemetry is a viable alternative to usual care for patients with T2DM and can lead to improvements in a wide range of outcomes. The inclusion of evidence from different study designs, such as reviews and trials, in our review strengthens the conclusion that use of telemetric interventions can be feasible in a clinical setting. Other reviews have also recently presented an improvement of clinical outcomes through telemetry and especially a trend toward a reduction in HbAc levels ,79. Our esources . In the 1c. All SRs and MAs also clearly showed that telemetric interventions improve HbA1c specifically, as well as the management of T2DM generally. Furthermore, “real-time video” interventions with a duration of 6 months were the most effective in reducing HbA1c. These interventions showed clear improvements in HbA1c levels in patients diagnosed with T2DM compared to usual care . Overall, the effects in the subgroup analysis in terms of the improvement of HbA1c values had MDs between –1.15% and –0.25%. These obvious decreases in HbA1c may indicate a novel and additional approach to diabetes care since these therapeutic effects could be accomplished by telemetric intervention alone. However, to optimize glucose homeostasis, individual telemetric approaches may be considered in terms of individual diabetes care as an addition to established therapeutic approaches [In general, all types of telemetric interventions clearly improved HbAThrough the use of “combined” interventions, FBG levels improved effectively, which was shown by a moderate number of studies. With “asynchronous” and “real-time audio” interventions, few studies showed an improvement in FBG values. However, the data were inconsistent as to whether “real-time video” interventions reduce FBG effectively.BP measurements decreased by applying “combined” interventions in a moderate number of the reviewed studies. “Asynchronous” and “real-time audio” interventions also improved BP, but there were comparatively few examinations. Moreover, the study situation for “real-time video” interventions was found to be rather inconsistent.Body weight decreased in a moderate number of studies by using “combined” interventions effectively. “Real-time audio” interventions also clearly reduced body weight in a few investigations. However, the study situation for “real-time video” interventions was not consistent.BMI decreased effectively in several studies by using “combined” interventions. The few studies available indicated that “asynchronous” and “real-time audio” interventions decreased BMI. In contrast, the study situation for “real time video” was inconsistent.Only “combined” interventions showed effective improvements regarding DRQoL and HRQoL, but there were few studies that examined DRQoL and very few studies of HRQoL compared with the other clinical outcomes. “Real-time audio,” “asynchronous,” and “combined” interventions were potentially cost-effective, but there was only a small number of studies. In addition, “real-time video,” “asynchronous,” and “combined” interventions occasionally showed time savings, although again, few studies examined these outcomes.1c clearly and effectively in short-term and long-term ways in a large number of studies. Weekly videoconferencing seems to be very effective in terms of reducing HbA1c. Due to the heterogeneity of the studies, the results regarding FBG, BP, body weight, BMI, and QoL may be rather inconsistent. However, they all have in common that user-friendly technologies were considered in the development of the interventions and that telemetry was anchored in people’s everyday lives, both of which are necessary for optimal results.“Real-time video” interventions did improve HbA1c, as demonstrated in numerous studies. Some studies indicated that there is also a clear beneficial impact of these interventions on FBG, BP, body weight, and BMI. Additionally, “real-time audio” interventions were shown to be cost-effective by the limited studies available.“Real-time audio” interventions proved to be effective in reducing HbA1c effectively. These interventions also improved FBG, BP, and BMI, and showed very positive results in terms of cost-effectiveness and time savings, but few studies using “asynchronous” interventions were available for review.Furthermore, a large number of studies pointed out that “asynchronous” interventions improved HbA1c values effectively. Furthermore, a moderate number of interventions had a favorable impact on FBG, BP, BMI, and body weight. In terms of DRQoL and HRQoL, there were few studies to examine these outcomes, but the available studies showed positive tendencies. Additionally, cost-effectiveness and time savings of telemetric interventions showed a positive trend, but sufficient data were lacking.Most studies assessed “combined” interventions . Numerous studies indicated that “combined” interventions improved HbAFrom our point of view, telemetric T2DM management enhances patient compliance, enables intensive monitoring, and empowers patients to deal with and understand their disease. For a successful implementation of telemetric approaches, it is also essential that the technology is user-friendly, that telemetric T2DM management can be easily integrated into everyday life, and that it is tailored to the patient and his or her life circumstances .Furthermore, we would like to point out that telemetric interventions differ not only in terms of their technologies but also in terms of their contextual focus  and that this aspect should be taken into account when interpreting the results.Although the exclusion criteria were observed, the included studies displayed a wide variation in terms of study design, technical and interventional approaches, duration, and frequency of contact with health care providers (in both the intervention and the control groups), as well as sample size and statistical evaluations used. Due to this heterogeneity, as well as to the small size of our MA, there may be potential for bias. For the same reasons, methodological quality and statistical evaluations could not be carried out. Some studies achieved improvements that were significant within the intervention groups but not between the groups, and methodological weaknesses may have been responsible for that.1c values (P<.05) from the implementation of telemetric interventions [1c. According to a review and network MA by Lee et al [1c by a mean of 0.43% (95% CI −0.64% to −0.21%). The authors concluded that all telemedical strategies, with the exceptions of telecase management and telementoring, were effective in reducing HbA1c in a clinically meaningful way. Furthermore, Mushcab et al [1c levels and quality of life. They also observed a high acceptance of web-based systems.Other research groups have displayed similar results. Numerous other SRs and MAs, which were included in this review, reported significant decreases in HbAlues P<.0 from theventions -17. Su eventions  examinedventions , who incee et al , over a Our research builds on these previous findings, incorporating a large number of studies (n=99), patients , and patient cases  and considering a range of main clinical outcomes in terms of T2DM management. Interestingly, there may be differences in telemetric approaches in terms of T2DM versus type 1 diabetes mellitus management , but theTo our knowledge, this is the first systematic meta-review analyzing telemetric approaches in T2DM management, including a wide range of important clinical outcomes and technologies.1c values in the short term and long term specifically and T2DM care generally. Moreover, “real-time video” interventions with a duration of 6 months showed the greatest effect in terms of improving HbA1c values in a sustained way. “Combined” interventions  appeared to be most effective in improving FBG, BP, body weight, BMI, and quality of life.Viewed together, telemetric interventions clearly improve HbA1c and T2DM management effectively. More studies need to be done, especially with a focus on main clinical outcomes.In conclusion, telemetric interventions clearly improve HbA