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Chlorella vulgaris is of great importance in numerous exploratory or industrial applications (e.g., medicals, food, and feed additives). Rapid quantification of algal biomass is crucial in photobioreactors for the optimization of nutrient management and the estimation of production. The main goal of this study is to provide a simple, rapid, and not-resource-intensive estimation method for determining the algal density of C. vulgaris according to the measured parameters using UV–Vis spectrophotometry. Comparative assessment measurements were conducted with seven different methods (e.g., filtration, evaporation, chlorophyll a extraction, and detection of optical density and fluorescence) to determine algal biomass. By analyzing the entire spectra of diluted algae samples, optimal wavelengths were determined through a stepwise series of linear regression analyses by a novel correlation scanning method, facilitating accurate parameter estimation. Nonlinear formulas for spectrometry-based estimation processes were derived for each parameter. As a result, a general formula for biomass concentration estimation was developed, with recommendations for suitable measuring devices based on algae concentration levels. New values for magnesium content and the average single-cell weight of C. vulgaris were established, in addition to the development of a rapid, semiautomated cell counting method, improving efficiency and accuracy in algae quantification for cultivation and biotechnology applications.

Background: Fabry disease is a progressive, X chromosome-linked lysosomal storage disorder with multiple organ dysfunction. Due to the absence or reduced activity of alpha-galactosidase A (AGAL), glycosphingolipids, primarily globotriaosyl-ceramide (Gb3), concentrate in cells. In heterozygous women, symptomatology is heterogenous and currently routinely used fluorometry-based assays measuring mean activity mostly fail to uncover AGAL dysfunction. The aim was the development of a flow cytometry assay to measure AGAL activity in individual cells. Methods: Conventional and multispectral imaging flow cytometry was used to detect AGAL activity. Specificity was validated using the GLA knockout (KO) Jurkat cell line and AGAL inhibitor 1-deoxygalactonojirimycin. The GLA KO cell line was generated via CRISPR-Cas9-based transfection, validated with exome sequencing, gene expression and substrate accumulation. Results: Flow cytometric detection of specific AGAL activity is feasible with fluorescently labelled Gb3. In the case of Jurkat cells, a substrate concentration of 2.83 nmol/mL and 6 h of incubation are required. Quenching of the aspecific exofacial binding of Gb3 with 20% trypan blue solution is necessary for the specific detection of lysosomal substrate accumulation. Conclusion: A flow cytometry-based assay was developed for the quantitative detection of AGAL activity at the single-cell level, which may contribute to the diagnosis of Fabry patients.

Abstract Background Imprinting disorders (ImpDis) comprise diseases which are caused by aberrant regulation of monoallelically and parent-of-origin-dependent expressed genes. A characteristic molecular change in ImpDis patients is aberrant methylation signatures at disease-specific loci, without an obvious DNA change at the specific differentially methylated region (DMR). However, there is a growing number of reports on multilocus imprinting disturbances (MLIDs), i.e. aberrant methylation at different DMRs in the same patient. These MLIDs account for a significant number of patients with specific ImpDis, and several reports indicate a central role of pathogenic maternal effect variants in their aetiology by affecting the maturation of the oocyte and the early embryo. Though several studies on the prevalence and the molecular causes of MLID have been conducted, homogeneous datasets comprising both genomic and methylation data are still lacking. Results Based on a cohort of 36 MLID patients, we here present both methylation data obtained from next-generation sequencing (NGS, ImprintSeq) approaches and whole-exome sequencing (WES). The compilation of methylation data did not reveal a disease-specific MLID episignature, and a predisposition for the phenotypic modification was not obvious as well. In fact, this lack of epigenotype–phenotype correlation might be related to the mosaic distribution of imprinting defects and their functional relevance in specific tissues. Conclusions Due to the higher sensitivity of NGS-based approaches, we suggest that ImprintSeq might be offered at reference centres in case of ImpDis patients with unusual phenotypes but MLID negative by conventional tests. By WES, additional MLID causes than the already known maternal effect variants could not be identified, neither in the patients nor in the maternal exomes. In cases with negative WES results, it is currently unclear to what extent either environmental factors or undetected genetic variants contribute to MLID.

Abstract Background Neurodevelopmental disorders are genetically heterogeneous pediatric conditions. The first tier diagnostic method for uncovering copy number variations (CNVs), one of the most common genetic etiologies in affected individuals, is chromosomal microarray (CMA). However, this methodology is not yet a routine molecular cytogenetic test in many parts of the world, including Hungary. Here we report clinical and genetic data of the first, relatively large Hungarian cohort of patients whose genetic testing included CMA. Methods Clinical data were retrospectively collected for 78 children who were analyzed using various CMA platforms. Phenotypes of patients with disease-causing variants were compared to patients with negative results using the chi squared/Fisher exact tests. Results A total of 30 pathogenic CNVs were identified in 29 patients (37.2%). Postnatal growth delay (p = 0.05564), pectus excavatum (p = 0.07484), brain imaging abnormalities (p = 0.07848), global developmental delay (p = 0.08070) and macrocephaly (p = 0.08919) were more likely to be associated with disease-causing CNVs. Conclusion Our results allow phenotypic expansion of 14q11.2 microdeletions encompassing SUPT16H and CHD8 genes. Variants of unknown significance (n = 24) were found in 17 patients. We provide detailed phenotypic and genetic data of these individuals to facilitate future classification efforts, and spotlight two patients with potentially pathogenic alterations. Our results contribute to unraveling the diagnostic value of rare CNVs.

Abstract Background Silver-Russell syndrome (SRS) is a genetic disorder characterized by intrauterine and postnatal growth restriction, relative macrocephaly at birth, body asymmetry and typical facial features. Clinical and molecular heterogeneity is described in SRS. Common causes are loss of methylation of the imprinting center 1 in 11p15 and maternal uniparental disomy of chromosome 7. Other genetic alterations include disturbances of imprinted regions in 14q32, 7q32 and 11p15 as well as submicroscopic deletions and duplications. Single nucleotide variants in genes like IGF2, HMGA2, PLAG1, CDKN1C have also been identified in patients with SRS phenotypes. However, routine molecular diagnostics usually focus on 11p15 and chromosome 7, while less frequent causes are not systematically addressed. Results Here we report two patients with SRS features in which molecular karyotyping revealed microdeletions in 1q21 and 8q12.1 respectively. In a 3.5-year-old girl with postnatal growth restriction, feeding difficulties, relative macrocephaly and distinct SRS features a 2 Mb deletion in 1q21.1q21.2 was identified. Our second case is a 1.5-year-old boy with intrauterine and postnatal growth restriction, feeding difficulties and distinct facial features with a 77 kb deletion in 8q12.1 affecting PLAG1 as the only protein-encoding gene with known function. Conclusions The 1q21 region has not yet been assigned as an SRS region, although six patients with the same deletion and SRS features including relative macrocephaly have been described before. This new case adds to the evidence that distal 1q21 should be annotated as an SRS candidate region. The PLAGL1 alteration is the smallest deletion in 8q12.1 ever reported in a patient with SRS phenotype and it finally confirms that PLAG1 is the SRS causing gene in 8q12.1. To increase the diagnostic yield in patients with suspected SRS, we recommend both molecular karyotyping and next generation sequencing-based approaches.

Fabry disease (FD) is a multisystemic X-linked lysosomal storage disease that presents with angiokeratomas (AKs). Our objective was to investigate the clinical and morphologic features of AKs and to present two experimental techniques, multispectral imaging (MSI) and non-linear microscopy (NLM). A thorough dermatological examination was carried out in our 26 FD patients and dermoscopic images (n = 136) were evaluated for specific structures. MSI was used for the evaluation of AKs in seven patients. NLM was carried out to obtain histology samples of two AKs and two hemangiomas. Although AKs were the most common manifestation, the majority of patients presented an atypical distribution and appearance, which could cause a diagnostic challenge. Dermoscopy revealed lacunae (65%) and dotted vessels (56%) as the most common structures, with a whitish veil present in only 25%. Autofluorescence (405 nm) and diffuse reflectance (526 nm) images showed the underlying vasculature more prominently compared to dermoscopy. Using NLM, AKs and hemangiomas could be distinguished based on morphologic features. The clinical heterogeneity of FD can result in a diagnostic delay. Although AKs are often the first sign of FD, their presentation is diverse. A thorough dermatological examination and the evaluation of other cutaneous signs are essential for the early diagnosis of FD.

Bioplastics have emerged as a promising alternative to conventional plastics, marketed as environmentally friendly and sustainable materials. They provide a variety of methods for efficient waste management contributing to the goals of the circular economy. At their end-of-life stage, bioplastics can generate added value through aerobic and anaerobic biological treatments (composting or anaerobic digestion). In this study, biomethane potential (BMP) tests were carried out under mesophilic conditions on eight different catering biodegradable plastics available in the market and certified as being biodegradable under industrial composting conditions. Chemical analysis of the biodegradable plastics included elemental analysis, Fourier-transform infrared spectroscopy, and inductively coupled plasma–optical emission spectrometry. Key differences were observed in total solids (TS) and volatile solids (VS) contents between the studied biopolymer products. TS values ranged between 85.00 ± 0.26% (Product 8) and 99.16 ± 0.23% (Product 4), whereas VS content ranged between 64.57 ± 0.25 %wm (Product 6) and 99.14 ± 0.17 %wm (Product 4). Elemental analysis (elements C, H, N, S, and O) was used to estimate the theoretical methane production (ThBMP) of each product. The highest ThBMP (538.6 ± 8.7 NmL/gVS) was observed in Product 4 correlated with the highest C and H contents, while the lowest ThBMP (431.8 ± 6.1 NmL/gVS) was observed in Product 2. Significant differences were recorded between BMP values according to the chemical composition of the polymers. The average of BMP values ranged between 50.4 ± 2.1 NmL/gVS and 437.5 ± 1.0 NmL/gVS. Despite being characterized by the same composition (cellulose/cellulose derivatives and calcium carbonate), Products 2, 3, and 6 revealed significant differences in terms of TS, VS, ThBMP, and BMP. Furthermore, a significant statistical relationship (p < 0.001) was found between time (days) and BMP values of the eight products (R2 = 0.899–0.964) during the initial phase. The study confirmed that cellulose-based materials can convert efficiently under mesophilic conditions into methane, at a relatively short retention time; hence, they can be regarded as a promising material for co-digestion with feedstock in industrial anaerobic biogas plants. In contrast, biodegradation of polylactic acids (PLA) does not occur under mesophilic conditions, and hence, pre-treatment of the polymers is recommended. Moreover, PLA-containing products are highly affected by the presence of other components (e.g., polybutylene adipate terephthalate and cellulose/cellulose derivatives).

There is a deep need to navigate within our genomic data to find, understand and pave the way for disease-specific treatments, as the clinical diagnostic journey provides only limited guidance. The human genome is enclosed in every nucleated cell, and yet at the single-cell resolution many unanswered questions remain, as most of the sequencing techniques use a bulk approach. Therefore, heterogeneity, mosaicism and many complex structural variants remain partially uncovered. As a conceptual approach, nanopore-based sequencing holds the promise of being a single-molecule-based, long-read and high-resolution technique, with the ability of uncovering the nucleic acid sequence and methylation almost in real time. A key limiting factor of current clinical genetics is the deciphering of key disease-causing genomic sequences. As the technological revolution is expanding regarding genetic data, the interpretation of genotype–phenotype correlations should be made with fine caution, as more and more evidence points toward the presence of more than one pathogenic variant acting together as a result of intergenic interplay in the background of a certain phenotype observed in a patient. This is in conjunction with the observation that many inheritable disorders manifest in a phenotypic spectrum, even in an intra-familial way. In the present review, we summarized the relevant data on nanopore sequencing regarding clinical genomics as well as highlighted the importance and content of pre-test and post-test genetic counselling, yielding a complex approach to phenotype-driven molecular diagnosis. This should significantly lower the time-to-right diagnosis as well lower the time required to complete a currently incomplete genotype–phenotype axis, which will boost the chance of establishing a new actionable diagnosis followed by therapeutical approach.

Biochars as soil amendments have been reported to improve soil properties and may have an important role in the mitigation of the consequences of climate change. As a novel approach, this study examines whether biochar and digestate co-application can be utilized as cost-effective, renewable plant nutrients. The effects of two types of biochar—wood chip biochar (WBC) and animal bone biochar (ABC), applied alone or in combination with an anaerobic digestate—on soil physicochemical properties, on the levels of selected elements, and on growth yields of ryegrass were studied in laboratory experiments. Most parameters were significantly affected by the treatments, and the investigated factors (biochar type, application rate, and the presence of digestate), as well as their interactions, were found to have significant effects on the characteristics investigated. The easily soluble phosphorus content (AL-P2O5) of the soil increased in all WBC and ABC biochar treatments, and the presence of digestate caused a further increase in AL-P2O5 in the case of anaerobic digestate-supplemented ABC treatment (ABCxAD). The pH increased in both ABC and WBC treatments, and also in the case of ABCxAD treatments. Similar increases in the salt content were detected in ABC-treated samples and in ABCxAD treatments at higher application rates. WBC increased the water holding capacity and carbon content of the soil. Phytotoxic effects of biochars were not detected, although higher doses resulted in slower germination. Combined biochar–digestate applications resulted in increased plant yields compared to sole biochar treatments. Thus, biochar–digestate combinations appear to be applicable as organo-mineral fertilizers.

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The aim of this study is to obtain a more complete picture of blood plasma melatonin concentrations in the donkey mares. To this purpose, sampling and statistical processing were carried out in such a way that allowed the researchers to establish the annual and daily rhythms. Based on human observations, according to the hypothesis of the authors, the blood plasma melatonin concentration of pregnant individuals rises during the late gestational period, before parturition. To confirm this, the melatonin concentrations of pregnant and non-pregnant jennies were monitored and compared. In regard to the circannual rhythm, the significantly lowest midnight melatonin value (27.67 pg mL–1) was typical for the summer solstice. Under consideration of circadian changes, a significantly strongest melatonin production (45.16 pg mL–1) was observed on the night of the winter solstice (p < 0.001). Considering gestational age, the blood plasma melatonin concentration (around 38 pg mL–1) does not change as gestation progresses (p = 0.136). The results obtained in this studied population of the domestic ass usefully expand the little knowledge previously gathered about the development of the blood plasma melatonin concentrations of this species.

This study investigates Thoroughbred mares exposed or not to supplemental blue light at the end of the gestation. Sixty mares and their 60 foals were selected for the investigation. Guard hair samples were collected from the shoulder just after (within 12 h) the parturition or birth. The foals of the light-treated mares developed significantly (p < 0.05) shorter hair than those of the control mares. A general effect of light treatment on basal hair diameter thinning could be demonstrated (p < 0.005). The maximum force of hair samples of light-treated mares and foals (0.098 and 0.085 N, respectively) was significantly lower than that of the control (0.272 and 0.178 N, respectively). The tensile strength (82.2 N/mm2) of the foal hair samples of the light-treated mares was significantly lower than that of the control foals (121.6 N/mm2). Although no significant difference was found in the elongation (ΔL), the hair of the control animals (mares and foals together) was more elastic than that of the treated animals (335 vs. 262 μm). In conclusion, the supplemental blue light treatment of the pregnant mares has a decreasing effect on both mares and their foals on the mechanical properties of the hair, making it shorter, thinner, and weaker.