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At least 19 recordsLinked to original sources

Adjusting batch effects in microarray expression data using empirical Bayes methods.

Non-biological experimental variation or "batch effects" are commonly observed across multiple batches of microarray experiments, often rendering the task of combining data from these batches difficult. The ability to combine microarray data sets is advantageous to researchers to increase statistical power to detect biological phenomena from studies where logistical considerations restrict sample size or in studies that require the sequential hybridization of arrays. In general, it is inappropriate to combine data sets without adjusting for batch effects. Methods have been proposed to filter batch effects from data, but these are often complicated and require large batch sizes ( > 25) to implement. Because the majority of microarray studies are conducted using much smaller sample sizes, existing methods are not sufficient. We propose parametric and non-parametric empirical Bayes frameworks for adjusting data for batch effects that is robust to outliers in small sample sizes and performs comparable to existing methods for large samples. We illustrate our methods using two example data sets and show that our methods are justifiable, easy to apply, and useful in practice. Software for our method is freely available at: http://biosun1.harvard.edu/complab/batch/.

Bayes Theorem↗

Moderated designs can balance between batch-effect mitigation and cell loss due to hashtag-assisted pooling in single-cell experiments.

Minimizing experimental noise is integral to robust data generation in single-cell omics. The current standard for avoiding batch effects during sample processing is barcode- or hashtag-assisted combining of different experimental treatments into one pool, allowing all samples to be subject to the technical protocols uniformly. The final data points for each treatment group are then computationally separated based on the original hashtag labels. Clearly, whereas hashtagging all groups and pooling them in a single well is expected to minimize batch effects, the procedure can also lead to a loss of cells that cannot be confidently decoded during the computational demultiplexing step. Here, we examine four alternate experimental designs, namely, compound, reference, chain, and confounded, that could be used instead of a single-pool approach and quantify the batch effects as well as cell loss in each case. We find a linear relationship-the percentage of cells lost is double the number of hashtags used in the experiment. We use these analyses to identify experimental designs that can successfully mitigate batch effects while minimizing multiplexing, hence the cell loss, in each well. Although a reference design offers the best overall performance, this study can help individual investigators choose particular approaches that are best suited for their biological questions.

Journal Article↗

NPM: latent batch effects correction of omics data by nearest-pair matching.

MOTIVATION: Batch effects (BEs) are a predominant source of noise in omics data and often mask real biological signals. BEs remain common in existing datasets. Current methods for BE correction mostly rely on specific assumptions or complex models, and may not detect and adjust BEs adequately, impacting downstream analysis and discovery power. To address these challenges we developed NPM, a nearest-neighbor matching-based method that adjusts BEs and may outperform other methods in a wide range of datasets. RESULTS: We assessed distinct metrics and graphical readouts, and compared our method to commonly used BE correction methods. NPM demonstrates the ability in correcting for BEs, while preserving biological differences. It may outperform other methods based on multiple metrics. Altogether, NPM proves to be a valuable BE correction approach to maximize discovery in biomedical research, with applicability in clinical research where latent BEs are often dominant. AVAILABILITY AND IMPLEMENTATION: NPM is freely available on GitHub (https://github.com/bigomics/NPM) and on Omics Playground (https://bigomics.ch/omics-playground). Computer codes for analyses are available at (https://github.com/bigomics/NPM). The datasets underlying this article are the following: GSE120099, GSE82177, GSE162760, GSE171343, GSE153380, GSE163214, GSE182440, GSE163857, GSE117970, GSE173078, and GSE10846. All these datasets are publicly available and can be freely accessed on the Gene Expression Omnibus repository.

Humans↗

Incorporating batch effects in the estimation of drug stability parameters using an Arrhenius model.

The nonlinear estimation of drug stability parameters (energy of activation Ea and shelf-life tY) by conventional approaches employs equations relating drug content determination C at time t and temperature T. The identification procedures lead to the determination of only one initial drug content C0 for several different experiments. However, it is well known that because of experimental concentration variation or of intentional modification of the experimental schedule, there are as many initial drug contents as experiments. For these reasons, a method which takes into account batch effects is proposed to determine stability parameters and also all initial drug contents C0j where j is the index of experiment in one step. This method is more accurate from a statistical viewpoint and is suitable for data treatment in pharmaceutical industries where the initial drug content of each batch entering the stability program can be checked a posteriori. The application of this method is shown on real kinetic data from the hydrolysis of acetylsalicylic acid (ASA).

Algorithms↗

Batch effects, water content and aqueous/organic solvent reactivity of microcrystalline cellulose samples.

The structural, morphological and surface features on two MCC powders of the same commercial type (Avicel PH 102), but coming from different countries (The Netherlands and Hong Kong) and vendors (DMV International and Mingtai Chemical Co., Ltd., respectively), have been investigated and compared, by means of the X-ray diffraction, SEM and BET and polymerization degree determination. TGA and water sorption from saturated vapor experiments have been applied to characterize and compare the MCC/water interactions of the two samples. The results were integrated by studies of preferential sorption from binary aqueous/organic solvents.

Acetonitriles↗

Genome- and peak-informed two-stage framework for scATAC-seq cell type identification.

MOTIVATION: Accurate cell type annotation is essential in scATAC-seq analysis, as it underpins the characterization of cellular heterogeneity, the identification of regulatory elements, and downstream biological discovery. However, current annotation methods still face major challenges. First, although some approaches attempt to integrate genomic sequence information, they typically rely on shallow sequence representations and thus fail to capture the long-range dependencies and regulatory signals encoded in DNA. Second, substantial batch effects introduced by different platforms, sequencing batches, or tissue sources remain insufficiently addressed. Existing models often lack robust distribution alignment and domain generalization capabilities, leading to confounding non-biological variation and reduced annotation accuracy across datasets. RESULTS: To overcome these limitations, we propose seqAlignATAC, a two-stage intra-modality annotation framework that integrates sequence-derived embeddings with domain adaptation. In the first stage, we employ a large-scale pretrained nucleotide language model to extract low-dimensional, biologically informative representations from the genomic sequences of chromatin-accessible peaks. In the second stage, these embeddings are fed into a supervised neural network equipped with an adaptive alignment module to mitigate batch effects and harmonize feature distributions between labeled reference and unlabeled target datasets. Extensive experiments across multiple settings demonstrate that seqAlignATAC achieves competitive accuracy and robustness, effectively leveraging genome-level information while alleviating batch-induced distributional discrepancies. AVAILABILITY AND IMPLEMENTATION: The source code of seqAlignATAC is available at: https://github.com/BioCS-Lab/seqAlignATAC.

Humans↗

[Animal model and multiple trait BLUP applied in poultry genetic evaluation].

Best linear unbiased prediction (BLUP) is a powerful method to estimate genetic values of animals, and is widely applied in many animal species but poultry. Beijing White Leghorn nested data in 1986-1987 with 777 individuals were analysed by animal model and multiple trait BLUP. Two traits (40-week egg production and 36-week egg weight) and two fixed effects (house-pen effect and hatching batch effect) were considered. The way to calculate a large set of mixed model equations in micro-computer was studied. Only non-zero elements of the coefficient matrix of MME were stored on a disk. The iteration process was reduced by block iteration. It also simplified the multiple trait BLUP method, the dimension of equations is only 1/q (q is the number of traits) of regular method. So it saved a lot of compute time and cost, and BLUP became applicable in poultry. The significance of using BLUP in poultry are: (1) Eliminating some fixed effects; (2) Reducing the estimation error for unbalanced data; (3) Estimating breeding values of progeny, so we can shorten the generation interval; (4) It can estimate breeding values of individuals without records from the relatives' information; (5) The breeding value of sires and dams can be estimated from their progeny records, used for family selection; (6) Due to genetic and environmental correlation between traits and all relatives information were considered, it can increase the selection accuracy.

Animals↗

H(2) production through anaerobic mixed culture: effect of batch S(0)/X(0) and shock loading in CSTR.

Biological production of H(2) has received considerable attention lately. The present study was undertaken to observe the effects of substrate/seeding ratios (S(0)/X(0)) on batch H(2) generation. The H(2)-producing seeding spores were obtained from the heat treatment (88 degrees C for 12h) of the compost from a grass composting facility. A dehydrated brewery mixture was used as feed substrate. The results indicate that the pattern of the cumulative H(2) production with time is similar to the growth curve with a typical lag, exponential and stationary phase; the results were successfully modeled with a modified Gompertz equation. It appears that maximum H(2) yield potential (27ml g(-1)COD(added)) occurs at an S(0)/X(0) ratio of about 4, whereas the maximum specific H(2) yield (205ml g(-1) VSSd(-1)) occurs at approximately S(0)/X(0)=3. The S(0)/X(0) ratios higher than 4 would inhibit H(2) production. An attempt was made to waste a certain amount of reactor content and replaced it with fresh substrate in order to enhance H(2) production. After this medium replacement, the H(2) production was initially inhibited and the system then exhibited a long lag before it reached an active H(2) production stage. For a continuous-stirred tank-reactor (CSTR) system, the results of replacing 25% of the reactor content indicate that there is still a lag time before a sudden increase in H(2) production after the addition of the new substrate feed. The major low molecular weight acids identified are HAc and HBu with total volatile acids of about 6000-8000mg l(-1). The ratio of HAc/HBu in the present study is relatively constant (about 5) and appears not significantly affected by the medium replacement. The concentration of total alcohols is about 2000mg l(-1). All in all, the CSTR system is able to recover to its previous performance after such a dramatic 25% medium replacement.

Bacteria↗

Whole metagenome sequencing: not deep enough for complete microbial function recovery.

BACKGROUND: Whole metagenome shotgun sequencing (WMS) is widely used to profile microbial function. However, technical variability in sequencing and analysis often obscures true biological patterns. Large-scale studies are particularly susceptible to batch effects, such as differences in sequencing depth and platform and annotation strategies, as well as sample-to-flow-cell assignments. However, the relative effects of these factors on functional inference in such studies have yet to be systematically evaluated. We analyzed oral-rinse WMS data from 671 Nigerian youths aged 9-18, sequenced on two Illumina platforms. Microbial molecular functionality encoded in these data was annotated using the mi-faser/Fusion pipeline, to capture the broad functional repertoire, and HUMAnN 3/EC numbers pipeline to characterize curated enzymatic activities. We then quantified how technical factors and batch effects shaped the recovery of microbial functionality. RESULTS: Three findings of our work were most salient. First, we observed that the choice of annotation strategy traded off between breadth and specificity of functional coverage. Second, we found that low-prevalence functions were disproportionately lost at shallow sequencing depths, indicating that in, e.g., case-control studies with few representatives of the minor class, sequencing depth could critically impact study resolution. Finally, using our newly developed model relating sequencing depth to functional recovery, we demonstrated that increasing sequencing depth does not directly or proportionally improve functional recall. That is, at as little as 10% of this study's sequencing depth, 30% of the estimated complete microbiome functional repertoire was detectable. However, even at the full depth used in this study, we were only able to recover an estimated 60% of that complete functional repertoire. We further showed that despite biomes differences in functional diversity and host contamination levels (e.g., soil, fecal), incomplete functional recovery at commonly used sequencing depths was consistently observed. CONCLUSIONS: Together, these findings and our depth-to-function mapping framework provide practical guidelines for the design and interpretation of WMS studies. Coordinating sequencing depth planning with annotation strategy, experimental design, and rigorous batch control is thus essential for robust detection of microbial functions and for ensuring reproducible microbiome insights. Video Abstract.

Humans↗

Aerobic biodegradation of 3-chlorophenol in a sequencing batch reactor: effect of cometabolism.

The aim of the present study was to investigate how phenol modifies, through cometabolism, the biodegrading capability of 3-chlorophenol (3-CP) in a sequencing batch reactor seeded with a mixed culture obtained from a domestic sewage treatment plant. Two laboratory-scale SBRs, one fed 3-CP only and the other fed 3-CP and phenol in the same concentration, were seeded with the partially acclimated biomass. The removal capability in both reactors was measured for progressive increases in the feed organic loading. Cometabolism enhanced biodegradation of 3-CP by reducing both the initial lag period and the time required for the complete removal. 700 mg/L 3-CP was demonstrated to be the highest concentration, which could be completely degraded during the active phase (fill plus react) either in the presence or absence of phenol as the growth substrate even though the lag period was shorter when phenol was present. The operating strategy required modification for the complete removal of 800 mg/L 3-CP. An increase in the phenol to 3-CP ratio did, however, improve 3-CP degradation rate.

Animals↗

Bayesian inference of genetic parameters and selection response for litter size components in pigs.

Three contemporary lines were formed from the progeny of 50 French Large White sows. In the first line, gilts were selected for ovulation rate at puberty. In the second line, they were selected for prenatal survival of the first two parities, corrected for ovulation rate. The control constituted the third line. Ovulation rate at puberty was analyzed using an animal model with a batch effect. Prenatal survival was analyzed with a repeatability animal model that included batch and parity effects. Flat priors were used to represent vague previous knowledge about parity and batch effects. Additive and residual effects were represented assuming that they were a priori normally distributed. Variance components were assumed to follow either uniform or inverted chi-square distributions, a priori. The use of different priors did not affect the results substantially. Heritabilities for ovulation rate ranged from 0.32 to 0.39, and from 0.11 to 0.16 for prenatal survival, depending on the prior used. The mean of the marginal posterior distribution of response to four generations of selection ranged from 0.38 to 0.40 ova per generation, and from 1.1 to 1.3% of the mean survival rate for average survival per generation.

Animals↗

Potential of two-dimensional electrophoresis in routine identification of closely related durum wheat lines.

Four closely related durum wheat varieties were compared by computer-assisted analysis of two-dimensional electrophoretic maps of leaf proteins. A low inter-varietal polymorphism was revealed and seven reliable qualitatively varying proteins allowed rapid visual identification of genotypes. For numerous spots, presence/absence or quantitative variations were greatly affected by a batch effect. Several criteria that should be used to discard unreliable spots or gels a priori were reviewed. Nevertheless, it was shown that, provided that the experimental design allows the integration of the batch effect, screening for discriminant markers as well as computing distances based on protein quantity variations are possible and allow variety identification. Euclidean and Mahalanobis distances allowed variety discrimination and single gel classification with a minimum risk of error, not only by taking into account the quantitative variations in discriminant proteins selected by analysis of variance, but also by taking into account all reproducible spots. The possible applications of two-dimensional electrophoresis in variety identification are discussed.

Autoanalysis↗

Enhanced biological phosphorus removal in an anaerobic-aerobic sequencing batch reactor: effect of pH.

Enhanced biological phosphorus removal (EBPR) is not always successfully achieved in activated-sludge processes by anaerobic-aerobic operation. It has been reported that the deterioration of EBPR has been caused by domination of glycogen-accumulating organisms (GAOs) in microbial sludge. After repeated operation of a sequencing batch reactor (SBR) supplied with acetate as a sole carbon source, it was found that an operational variable, pH, could be a tool to control the competition between GAOs and polyphosphate-accumulating organisms PAOs, which could mediate the success of EBPR. When anaerobic phase pH in the SBR was controlled at 7.0, GAOs dominated the culture, resulting in failure of EBPR. Without pH control, the pH during the anaerobic phase increased to 8.4 as a result of denitrification and acetate uptake. In this relatively high pH condition, almost complete EBPR was achieved. In this report, the mechanism of pH effect on the competition between GAOs and PAOs was elucidated through a characterization of activated sludge using previously proposed biochemical models.

Acetates↗

Defined medium for Aquaspirillum serpens VHL effective in batch and continuous culture.

A defined medium for Aquaspirillum serpens VHL allows the replacement of the complex media now in use. It was developed by batch culture methods but supports growth in continuous culture. A basal salts medium supplemented with L-aspartic acid, L-alanine, and L-glutamic acid provided the best growth (turbidity), as long as ammonium chloride was omitted. Ammonium chloride caused either a lag or a reduction or a complete inhibition of the growth of A. serpens VHL on the above amino acids and other organic supplements depending on the combination used. Ammonium sulfate and ammonium hydroxide with L-glutamic acid allowed growth, but the lag period was increased in shake flask cultures. Vitamins, cysteine hydrochloride, and carbon dioxide had no effect on the growth rate. Viability (less than 50%) was inadequate to maintain continuous culture with L-glutamic acid as the sole source of carbon and nitrogen. Combinations of amino and carboxylic acids were then tested and, of these, L-glutamic acid (1 g/liter) and L-histidine (75 mg/liter) without ammonium chloride in the basal salts medium supported growth in batch and continuous culture. L-Glutamic acid was the limiting substrate for growth.

Alanine↗

Research on identification of key genes and immune-metabolic mechanisms in atrial fibrillation through integrated multi-cohort transcriptomic analysis and machine learning.

This study aimed to integrate multiple datasets for the identification of atrial fibrillation (AF)-related differentially expressed genes (DEGs), analyze their underlying mechanisms through functional enrichment and machine learning, construct diagnostic models, and explore immune-metabolic interactions to provide novel biomarkers and theoretical foundations. Gene expression datasets were integrated and normalized, with batch effects removed using principal component analysis. Differential expression analysis, functional enrichment analysis (Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways), and machine learning-based feature gene selection and model construction were performed. Shapley additive explanations analysis was utilized to interpret the constructed models, while gene set enrichment analysis, gene set variation analysis, and immune cell infiltration analysis were conducted to investigate the associations between feature genes and immune infiltration. After integrating and normalizing gene expression data and eliminating batch effects via principal component analysis, 6 DEGs were identified, including 4 upregulated and 2 down-regulated ones. Functional enrichment analysis showed these DEGs were significantly enriched in neuro-related biological processes and pathways, indicating their key roles in AF pathogenesis. Five key feature genes were selected using LASSO, random forest, and support vector machine-recursive feature elimination algorithms. They had significant expression differences between the AF and control groups (P&#x2005;<&#x2005;.001) and were located on distinct chromosomes. The constructed random forest and support vector machine models performed excellently (area under the curve&#x2005;&#x2265;&#x2005;0.85). Shapley additive explanations analysis revealed TNNI1 contributed most to model prediction, with its expression significantly positively correlated with immune cell infiltration. Gene set enrichment analysis and gene set variation analysis analyses further showed feature genes participated in AF pathogenesis by regulating immune modulation, metabolic pathways, and autophagy. Immune cell infiltration analysis found altered proportions of T-cell subsets and M0 macrophages in the AF group, along with complex links between feature gene expression and immune cell function. This study systematically elucidated the unique gene expression patterns and key regulatory pathways associated with AF, clarifying the crucial roles of feature genes in immune regulation, metabolic imbalance, and cellular dysfunction. These findings provide a theoretical basis and potential therapeutic targets for understanding AF pathogenesis and developing targeted treatment strategies.

Atrial Fibrillation↗

Effects of batch to batch variations and test methodology on degree of crystallinity and melting temperature of UHMW-PE as measured by differential scanning calorimetry.

The thermal and structural analysis of a total 22 batches from three medical grade ultrahigh molecular weight polyethylene (UHMW-PE) resins was determined using the DSC method. Batch to batch variations of approximately 5% were obtained for the melting temperature and the degree of crystallinity. The variations correlated both to the changes caused by gamma-irradiation sterilization and subsequent aging, and to the differences in DSC results obtained from various laboratories on materials believed to exhibit essentially similar clinical performances. Varying the sample weight from 4.4 to 27.1 mg resulted in a significant relative increase of 5% in the degree of crystallinity. The study shows that the standard double melting/recrystallisation procedure for DSC evaluation produces variations in the melting behaviors of approximately the same magnitude as those indicated by batch lot or sterilization/aging.

Calorimetry, Differential Scanning↗

Effects of batch variation on shade of dental porcelain.

Three different batches of three commercially available porcelains were applied to 36 specimens of two noble alloys. The shades were visually compared by nine observers. Results indicated the following. No color differences were found among specimens of individual batches. Discerning color changes were observed among three batches of one brand of porcelain. The porcelain fired in the laboratory may not match the manufacturer's standard shade guide. Batch to batch variation of porcelain may necessitate the fabrication of customized shade tabs with fresh batches of porcelain.

Chemical Phenomena↗