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The female reproductive system can face serious disorders and show reproductive abnormalities under the influence of environmental pollutants. Microplastics (MPs) and nanoplastics (NPs) as emerging pollutants, by affecting different components of this system, may make female fertility a serious challenge. Animal studies have demonstrated that exposure to these substances weakens the function of ovaries and causes a decrease in ovarian reserve capacity. Also, continuous exposure to micro/nano plastics (MNPs) leads to increased levels of reactive oxygen species, induction of oxidative stress, inflammatory responses, apoptosis of granulosa cells, and reduction of the number of ovarian follicles. Furthermore, by interfering with the hypothalamic-pituitary-ovarian axis, these particles disturb the normal levels of ovarian androgens and endocrine balance and delay the growth of gonads. Exposure to MNPs can accelerate carcinogenesis in the female reproductive system in humans and animal models. Animal studies have determined that these particles can accumulate in the placenta, causing metabolic changes, disrupting the development of the fetus, and endangering the health of future generations. In humans, the presence of micro/nanoplastics in placenta tissue, infant feces, and breast milk has been reported. These particles can directly affect the health of the mother and fetus, increasing the risk of premature birth and other pregnancy complications. This review aims to outline the hazardous effects of micro/nano plastics on female reproductive health and fetal growth and discuss the results of animal experiments and human research focusing on cellular and molecular pathways. [ABSTRACT FROM AUTHOR]

Research on knowledge distillation has become active in deep neural networks. Knowledge distillation involves training a low-capacity model from a high-capacity model. However, when the capacities of the teacher and student models differ, it can result to poor learning and low generalization performance. We propose here a novel teacher assistant model called Knowledge in Attention Assistant. This model learns learning a discriminative representation of important regions and statistical information along with spatial and channel knowledge. Moreover, by using a triplet attention mechanism, the student model can learn both the inner and outer distribution of different categories, and also memorize the knowledge distribution of the teacher model. This alignment improves the effectiveness and generalization of knowledge distillation and reduces the capacity gap between the teacher and student models. The present model addresses feature inconsistency by adjusting the attention weight distribution based on the resemblance between the features of the teacher and student. The evaluation of the proposed teacher assistant method shows remarkable results. The student model outperforms the teacher model in terms of generalization performance, achieving improvements of 93.37% and 94.09% on CIFAR-10 and CIFAR-100 datasets, respectively. Furthermore, the proposed model enhances the F1-scores 91.98% on CIFAR-10 and 79.69% on CIFAR-100. [ABSTRACT FROM AUTHOR]

Most deep neural networks (DNNs) are trained in an over-parametrized regime. In this case, the numbers of their parameters are more than available training data which reduces the generalization capability and performance on new and unseen samples. Generalization of DNNs has been improved by applying various methods such as regularization techniques, data enhancement, network capacity restriction, injection randomness, etc. In this paper, we proposed an effective generalization method, named multivariate statistical knowledge transformation, which learns feature distribution to separate samples based on the variance of deep hypothesis space in all dimensions. Moreover, the proposed method uses latent knowledge of the target to boost the confidence of its prediction. Compared to state-of-the-art methods, the transformation of multivariate statistical knowledge yields competitive results. Experimental results show that the proposed method achieved impressive generalization performance on CIFAR-10, CIFAR-100, and Tiny ImageNet with accuracy of 91.96%, 97.52%, and 99.21% respectively. Furthermore, this method enables faster convergence during the initial epochs. [ABSTRACT FROM AUTHOR]

Two modified ranked set sampling schemes are used to estimate the upper and lower quantiles of an underlying distribution. The performance of these sampling schemes is compared with that of ordinary ranked set sampling in view of Pitman's measure of closeness criterion. Actually, a way of choosing estimators is proposed in the paper based on Pitman closeness which demonstrates that the mentioned modifications on ranked set sampling are useful in the problem of quantile estimation. The results are applied to the location-scale family of distributions and the Pitman closeness probabilities are obtained numerically for the cases of exponential and uniform distributions. It is shown that the proposed sampling schemes would improve the performance of the point estimators of the population quantiles specially for extreme quantiles. The proposed procedure is used to estimate the quantiles of a real data set. [ABSTRACT FROM AUTHOR]

Introduction: Recent studies have acknowledged the potential of convolutional neural networks (CNNs) in distinguishing healthy and morbid samples by using heart sound analyses. Unfortunately the performance of CNNs is highly dependent on the filtering procedure which is applied to signal in their convolutional layer. The present study aimed to address this problem by applying filter bank learning concept in CNNs. Material and Methods: In proposed method, the filter bank of CNN is updated based on a cross-entropy minimization rule to extract higher-level features from spectral characteristics of the heart sound signal. The deeper level of the extracted features in parallel with their spectral-based nature leads to better discrimination between healthy and morbid heart sounds. The proposed method was applied to three different heart sound datasets of PASCAL-A, PASCAL-B, and Kaggle, including normal and abnormal categories. Results: The proposed method obtained a true positive rate (TPR) between minimally 86% and maximally 96% (if FPR=0%) among all the examined datasets. In addition, the false-positive rate (FPR) was obtained as 7-8% (if TPR=100%) among the mentioned datasets. Finally, the accuracy was achieved in the range of 93- 98% when the FPR was 0% and within the range of 96-96.5% when the TRP was 100%. Conclusion: Increased TPR in the proposed method (96% for the proposed method vs. 87% for CNN) in parallel with a decrease in its FPR (7% for the proposed method vs. 10% for CNN) showed the proposed method's superiority against its well-known alternative in automated self-assessment of the heart. [ABSTRACT FROM AUTHOR]

Background: P300 signal detection is an essential problem in many fields of Brain-Computer Interface (BCI) systems. Although deep neural networks have almost ubiquitously used in P300 detection, in such networks, increasing the number of dimensions leads to growth ratio of saddle points to local minimums. This phenomenon results in slow convergence in deep neural network. Hyperparameter tuning is one of the approaches in deep learning, which leads to fast convergence because of its ability to find better local minimums. In this paper, a new adaptive hyperparameter tuning method is proposed to improve training of Convolutional Neural Networks (CNNs). Methods: The aim of this paper is to introduce a novel method to improve the performance of deep neural networks in P300 signal detection. To reach this purpose, the proposed method transferred the non-convex error function of CNN) into Lagranging paradigm, then, Newton and dual active set techniques are utilized for hyperparameter tuning in order to minimize error of objective function in high dimensional space of CNN. Results: The proposed method was implemented on MATLAB 2017 package and its performance was evaluated on dataset of Ecole Polytechnique Fédérale de Lausanne (EPFL) BCI group. The obtained results depicted that the proposed method detected the P300 signals with 95.34% classification accuracy in parallel with high True Positive Rate (i.e., 92.9%) and low False Positive Rate (i.e., 0.77%). Conclusions: To estimate the performance of the proposed method, the achieved results were compared with the results of Naive Hyperparameter (NHP) tuning method. The comparisons depicted the superiority of the proposed method against its alternative, in such way that the best accuracy by using the proposed method was 6.44%, better than the accuracy of the alternative method. [ABSTRACT FROM AUTHOR]

The phosphoenolpyruvate-dependent phosphotransferase system (PTS) is the main carbohydrate uptake system in Bacillus subtilis. A typical PTS consists of two general proteins, enzyme I (EI) and a histidine-containing protein (HPr), as well as a specific carbohydrate transporter (or enzyme II [EII]), all of which transfer the phosphoryl group from phosphoenolpyruvate to the transported carbohydrate. The specific PTS transporters are formed by multidomain proteins or single-domain subunits. These domains are domain C (EIIC), the transmembrane channel for the carbohydrate transport; domain B (EIIB), the membrane-bound domain responsible for phosphorylation of the carbohydrate; and domain A (EIIA), the mediator between HPr(H15~P) and EIIB. There are 16 PTS transporters in B. subtilis, 6 of which, i.e., NagP, MalP, MurP, TreP, SacP, and SacX, contain no EIIA domain. Deletion of the single-EIIA-containing transporters showed that there is cross talk between the noncognate EIIA and EIIB domains in PTS. By deletion of all EIIA-containing proteins, strain KM455 (ΔEIIA) was constructed, and the EIIA-containing proteins were individually introduced into the strain. In this way, the PTS transporters of the glucose family, namely, PtsG, GamP, and PtsA (also known as YpqE), enabled growth with maltose, N-acetylglucosamine, sucrose, or trehalose as the sole carbon source. Construction of TkmA-EIIA fusion proteins confirmed the probable interaction between the EIIAs of the glucose family of PTS transporters and the EIIA-deficient PTS transporters. Likewise, we have shown that SacX is mainly phosphorylated by PtsA and GamP. PtsG and GmuA were also able to phosphorylate SacX, albeit less well than GamP and PtsA. [ABSTRACT FROM AUTHOR]

Bacillus subtilis possesses different enzymes for the utilization of plant cell wall polysaccharides. This includes a gene cluster containing galactan degradation genes (ganA and ganB), two transporter component genes (ganQ and ganP), and the sugarbinding lipoprotein-encoding gene ganS (previously known as cycB). These genes form an operon that is regulated by GanR. The degradation of galactan by B. subtilis begins with the activity of extracellular GanB. GanB is an endo-β-1,4-galactanase and is a member of glycoside hydrolase (GH) family 53. This enzyme was active on high-molecular-weight arabinose-free galactan and mainly produced galactotetraose as well as galactotriose and galactobiose. These galacto-oligosaccharides may enter the cell via the GanQP transmembrane proteins of the galactan ABC transporter. The specificity of the galactan ABC transporter depends on the sugar-binding lipoprotein, GanS. Purified GanS was shown to bind galactotetraose and galactotriose using thermal shift assay. The energy for this transport is provided by MsmX, an ATP-binding protein. The transported galacto-oligosaccharides are further degraded by GanA. GanA is a β-galactosidase that belongs to GH family 42. The GanA enzyme was able to hydrolyze short-chain β-1,4-galacto-oligosaccharides as well as synthetic β-galactopyranosides into galactose. Thermal shift assay as well as electrophoretic mobility shift assay demonstrated that galactobiose is the inducer of the galactan operon regulated by GanR. DNase I footprinting revealed that the GanR protein binds to an operator overlapping the -35 box of the σA-type promoter of Pgan, which is located upstream of ganS. [ABSTRACT FROM AUTHOR]

Consider a two-sampling scheme in which an initial sample is first taken from the underlying population and then by assuming a suitable restriction on this sample, some more data points are observed as a new restricted sample. This sampling scheme is used to do inference about the lower quantiles of the underlying distribution. The results are compared with those of simple random sampling in view of mean squared error and Pitman’s measure of closeness criteria for exponential and uniform distributions. It will be shown that the proposed sampling scheme would improve the performance of the point estimators of the lower quantiles of the population. [ABSTRACT FROM PUBLISHER]

Bacillus subtilis is a widely used bacterium for production of heterologous and homologous proteins. The primary challenge in the production of proteins in B. subtilis is choosing a relevant expression system. In this study, we developed a robust expression system based on optimized P tetR of transposon Tn 1721 , which is repressible by its specific repressor, TetR. The first step of this work was focused on the optimization of structure and core elements of Tn 1721 anhydrotetracycline-inducible promoters, P tetA and P tetR . Both promoters were inserted upstream of eGFP on a pUB110-derivative with high copy number. Reduction of the 18 bp spacer region of both P tetA and P tetR to 17 bp significantly increased their strength in B. subtilis . Nevertheless, only the optimized P tetR with 17 bp spacer region (P tetR 2 ) directed high level of eGFP expression. In the second step, regulation of the system was optimized by testing the expression of tetR using well-known promoters, such as P mtlA , P mtlR , P ptsG and P penP . Expression of tetR by P ptsG resulted in a tight regulation of P tetR 2 - eGFP showing 44-fold induction. By using the final expression plasmid in B. subtilis , neopullulanase was produced up to 15% of the total soluble protein. [ABSTRACT FROM AUTHOR]

Corynebacterium glutamicum is able to utilize vanillate, the product of lignin degradation, as the sole carbon source. The vanillate utilization components are encoded by the vanABK operon. The vanA and vanB genes encode the subunits of vanillate O-demethylase, converting vanillate to protocatechuate, while VanK is the specific vanillate transporter. The vanABK operon is regulated by a PadR-type repressor, VanR. Heterologous gene expression and variations of the vanR open reading frame revealed that the functional VanR contains 192 residues (21 kDa) and forms a dimer, as analyzed by size exclusion chromatography. In vivo, ferulate, vanillin, and vanillate induced PvanABK in C. glutamicum, while only vanillate induced the activity of PvanABK in Escherichia coli lacking the ferulate catabolic system. Differential scanning fluorimetry verified that vanillate is the only effector of VanR. Interaction between the PvanABK DNA fragment and the VanR protein had an equilibrium dissociation constant (KD) of 15.1 ± 1.7 nM. The VanR-DNA complex had a dissociation rate constant (Kd) of (267 ± 23) x 10-6 s-1, with a half-life of 43.5 ± 3.6 min. DNase I footprinting localized the VanR binding site at PvanABK, extending from +9 to +45 on the coding strand. Deletion of the nucleotides +18 to +27 inside the VanR binding site rendered PvanABK constitutive. Fusion of the T7 promoter and the wild-type VanR operator, as well as its shortened versions, indicated that the inverted repeat AACTAACTAA(N4)TTAGGTATTT is the specific VanR binding site. It is proposed that the VanR-DNA complex contains two VanR dimers at the VanR operator. [ABSTRACT FROM AUTHOR]

Objectives: Consumption of fruits, such as plums and prunes, is useful in treating blood circulation disorder, measles, digestive disorder, and prevention of cancer, diabetes, and obesity. The paper presents a description of antioxidant and antiradical capacity of plum (Prunus domestica L.) in both fresh and dried samples. Materials and Methods: Samples were mixed with methanol and ethanol (as solvents) and were extracted on magnetic shaker, separately. The experiments were carried out to measure the Total Phenolic Content (TPC), Total Flavonoid Content (TFC), Total Antioxidant Capacity (TAC), Reducing Power Assay (RPA), Chain Breaking Activity (CBA), and quantity of Malondialdehyde (MDA), 2,2-Diphenyl-1-Picrylhydrazyl (DPPH),Nitric Oxide (NO), Hydrogen peroxide (H2O2) and superoxide(O2-) radicals inhibition. Results: The results showed that the highest values for the TPC, TFC, TAC, RPA, CBA, DPPH, and NO were related to ethanolic extractsof dried sample which showed statistically significant differences (p<0.01 and p<0.0001), while the maximum values for the H2O2 and O2 -were related to ethanolic extracts of fresh sample. The correlations data were analyzed among all parameters and the TPC and TFC had a significant correlation (r2=0.977). Moreover, it was found that methanol was more successful in extraction procedure than ethanol (p<0.01). Conclusion: Findings suggest that the fresh samples are more successful in collecting oxygen free radicals such as superoxide (O2-) and peroxy radicals (ROO.) than dried. [ABSTRACT FROM AUTHOR]

• A new hydrological regions delineation method based on changepoint analysis is proposed. • Similarity between regions is based on the location of changepoints in time series. • The method allows the estimation of local and regional changing linear trends. • The method was applied to 11 precipitation series in the Tensift basin, Morocco. • Cross-wavelet analysis is used to confirm the homogeneity of the delineated regions. Hydrologic regionalization consists of regrouping stations and catchments in pools based on a similarity measure. Regionalization is commonly used to extract a robust signal that can be used to describe the hydrology of the region or extrapolated to a location without measured information. Obviously, the similarity measure used affects the type of hydrological behavior one would expect from stations within a region. Most regionalization methods assume a stable and/or linear relationship between parameters of interests while it is well known that the physical processes driving the behavior of hydrometeorological variables are inherently non-linear and non-stationary. In this paper, we propose a similarity measure that is based on the location of changepoints in hydrological time series. The proposed method has the unique advantage over other hydrological region delineation methods to detect regions where hydrological member stations are non-linearly correlated, and where the strength of the relation varies with time. It therefore has the potential to uncover similarities that would not have been detected by existing regionalization techniques. The proposed method is applied to the Tensift watershed located in Morocco, North Africa. The coherence of the detected regions is checked using wavelet coherence. [ABSTRACT FROM AUTHOR]

Bacillus subtilis is a heterotrophic soil bacterium that hydrolyzes different polysaccharides mainly found in the decomposed plants. These carbohydrates are mainly cellulose, hemicellulose, and the raffinose family of oligosaccharides (RFOs). RFOs are soluble a-galactosides, such as raffinose, stachyose, and verbascose, that rank second only after sucrose in abundance. Genome sequencing and transcriptome analysis of B. subtilis indicated the presence of a putative a-galactosidase-encoding gene (melA) located in the msmRE-amyDC-melA operon. Characterization of the MelA protein showed that it is a strictly Mn2+- and NAD+-dependent a-galactosidase able to hydrolyze melibiose, raffinose, and stachyose. Transcription of the msmER-amyDC-melA operon is under control of a sA-type promoter located upstream of msmR (PmsmR), which is negatively regulated by MsmR. The activity of PmsmR was induced in the presence of melibiose and raffinose. MsmR is a transcriptional repressor that binds to two binding sites at PmsmR located upstream of the -35 box and downstream of the transcriptional start site. MsmEX-AmyCD forms an ATP-binding cassette (ABC) transporter that probably transports melibiose into the cell. Since msmRE-amyDC-melA is a melibiose utilization system, we renamed the operon melREDCA. [ABSTRACT FROM AUTHOR]

Bacillus subtilis phosphorylates sugars during or after their transport into the cell. Perturbation in the conversion of intracellular phosphosugars to the central carbon metabolites and accumulation of phosphosugars can impose stress on the cells. In this study, we investigated the effect of phosphosugar stress on B. subtilis. Preliminary experiments indicated that the nonmetabolizable analogs of glucose were unable to impose stress on B. subtilis. In contrast, deletion of manA encoding mannose 6-phosphate isomerase (responsible for conversion of mannose 6-phosphate to fructose 6-phosphate) resulted in growth arrest and bulged cell shape in the medium containing mannose. Besides, an operon encoding a repressor (GlcR) and a haloic acid dehalogenase (HAD)-like phosphatase (PhoC; previously YwpJ) were upregulated. Integration of the PglcR-lacZ cassette into different mutational backgrounds indicated that PglcR is induced when (i) a manA-deficient strain is cultured with mannose or (ii) when glcR is deleted. GlcR repressed the transcription of glcR-phoC by binding to the σA-type core elements of PglcR. An electrophoretic mobility shift assay showed no interaction between mannose 6-phosphate (or other phosphosugars) and the GlcR-PglcR DNA complex. PhoC was an acid phosphatase mainly able to dephosphorylate glycerol 3-phosphate and ribose 5-phosphate. Mannose 6-phosphate was only weakly dephosphorylated by PhoC. Since deletion of glcR and phoC alone or in combination had no effect on the cells during phosphosugar stress, it is assumed that the derepression of glcR-phoC is a side effect of phosphosugar stress in B. subtilis. [ABSTRACT FROM AUTHOR]

The complex regulatory network in Bacillus, known as quorum sensing, offers many opportunities to modify bacterial gene expression and hence to control bioprocesses. One target regulated by this mechanism is the activity of the PsrfA promoter, which is engaged in the formation of lipopeptide surfactin. It was hypothesised that deletion of rapC, rapF and rapH, encoding for prominent Rap-phosphatases known to affect PsrfA activity, would enhance surfactin production. Therefore, these genes were deleted in a sfp+ derivative of B. subtilis 168 with subsequent evaluation of quantitative data. Up to the maximum product formation of the reference strain B. subtilis KM1016 after 16 h of cultivation, the titers of the rap deletion mutants did not exceed the reference. However, an increase in both product yield per biomass YP/X and specific surfactin productivity qsurfactin was observed, without any considerable effect on the ComX activity. By extending the cultivation time, a 2.7-fold increase in surfactin titer was observed after 24 h for strain CT10 (ΔrapC) and a 2.5-fold increase for CT11 (ΔrapF) compared to the reference strain KM1016. In addition, YP/X was again increased for strains CT10 and CT11, with values of 1.33 g/g and 1.13 g/g, respectively. Interestingly, the effect on surfactin titer in strain CT12 (ΔrapH) was not as distinct, although it achieved the highest promoter activity (PsrfA-lacZ). The data presented support the possibility of involving the quorum sensing system of Bacillus in bioprocess control as shown here on the example of lipopeptide production. [ABSTRACT FROM AUTHOR]

In today's competitive environment, taking the advantage of supply chain which is both effective and efficient is a competitive advantage. An adequate supply chain has a multifaceted structure that focuses on integrating all factors, including pricing. The final product is delivered to customers based on maintaining market values and increasing strategic interactions. Present study aims to present the most effective functioning of competitive supply chain pricing in accordance with aspects of social responsibility. In doing so, we conducted a case study on oil-related products of Iranian knowledge-based companies. A combined analysis with 15 university-level industrial management experts was employed in this study to establish the components (competitive supply chain pricing function) and research propositions (dimensions of social responsibility). Twenty managers of knowledge-based enterprises in the oil field evaluated the components and propositions identified in the form of matrix questions using interpretative rating (IRP). The results showed that the proposition of the ethical dimension is the most effective measure of the social responsibility of knowledge-based companies in the oil field, creating competitive supply chain pricing functions based on value. This result shows that, under the ethical dimension of social responsibility, processes of competitive supply chain pricing are formed in accordance with the customer values and, according to the strategic relationship between the company and the economy with regional and global partners, maintaining values are a critical factor in developing our interactions between pricing factors. Given the results, in the first step for success, knowledge-based companies are recommended to consider the priorities of professional ethics in addition to knowledge in order to be able to gain the trust of customers by understanding their functional nature in the country's developing economy. In this situation, the pricing process based on the creation of comprehensive values for customers can help to increase the company's competitive position and make these companies to be at the first level of strategic interactions with companies that use petroleum products. [ABSTRACT FROM AUTHOR]

In the modern era, intellectual capital encompasses all resources within an organization that enhance the value and competence of the organization. Consequently, this indicates that managing intellectual capital effectively will enhance the value and performance of an organization. This study aims to investigate the effects of intellectual capital on business performance through the use of customer knowledge management in the Bank Mellat branches of Iran. In this study, all managers and employees working for Bank Mellat in Tehran are included. Based on Morgan's table, the sample size was 220 people. Sampling was done by the simple random method. We used a descriptive correlation method to conduct this study and a questionnaire was used to collect data. The questionnaires were scored using a Likert scale. It was confirmed by a consensus of experts that the research instrument was valid, and the reliability of the research was 0.894%. Structural equation modeling was used to analyze the data. According to the results, the dimensions of intellectual capital (human, structural, and relational) have a significant impact on business performance. However, relational capital has been more influential on business performance than other factors. [ABSTRACT FROM AUTHOR]

Wild-type cultivations are of invaluable relevance for industrial biotechnology when it comes to the agricultural or food sector. Here, genetic engineering is hardly applicable due to legal barriers and consumer's demand for GMO-free products. An important pillar for wild-type cultivations displays the genus Bacillus. One of the challenges for Bacillus cultivations is the global ComX-dependent quorum sensing system. Here, molecular process control can serve as a tool to optimize the production process without genetic engineering. To realize this approach, quantitative knowledge of the mechanism is essential, which, however, is often available only to a limited extent. The presented work provides a case study based on the production of cyclic lipopeptide surfactin, whose expression is in dependence of ComX, using natural producer B. subtilis DSM 10 T. First, a surfactin reference process with 40 g/L of glucose was performed as batch fermentation in a pilot scale bioreactor system to gain novel insights into kinetic behavior of ComX in relation to surfactin production. Interestingly, the specific surfactin productivity did not increase linearly with ComX activity. The data were then used to derive a mathematic model for the time course of ComX in dependence of existing biomass, biomass growth as well as a putative ComX-specific protease. The newly adapted model was validated and transferred to other batch fermentations, employing 20 and 60 g/L glucose. The applied approach can serve as a model system for molecular process control strategies, which can thus be extended to other quorum sensing dependent wild-type cultivations. [ABSTRACT FROM AUTHOR]

Background: Bacillus subtilis is a well-established host for a variety of bioproduction processes, with much interest focused on the production of biosurfactants such as the cyclic lipopeptide surfactin. Surfactin production is tightly intertwined with quorum sensing and regulatory cell differentiation processes. As previous studies have shown, a non-sporulating B. subtilis strain 3NA encoding a functional sfp locus but mutations in the spo0A and abrB loci, called JABs32, exhibits noticeably increased surfactin production capabilities. In this work, the impacts of introducing JABs32 mutations in the genes spo0A, abrB and abh from 3NA into strain KM1016, a surfactin-forming derivative of B. subtilis 168, was investigated. This study aims to show these mutations are responsible for the surfactin producing performance of strain JABs32 in fed-batch bioreactor cultivations. Results: Single and double mutant strains of B. subtilis KM1016 were constructed encoding gene deletions of spo0A, abrB and homologous abh. Furthermore, an elongated abrB version, called abrB*, as described for JABs32 was integrated. Single and combinatory mutant strains were analysed in respect of growth behaviour, native PsrfA promoter expression and surfactin production. Deletion of spo0A led to increased growth rates with lowered surfactin titers, while deletion or elongation of abrB resulted in lowered growth rates and high surfactin yields, compared to KM1016. The double mutant strains B. subtilis KM1036 and KM1020 encoding Δspo0A abrB* and Δspo0A ΔabrB were compared to reference strain JABs32, with KM1036 exhibiting similar production parameters and impeded cell growth and surfactin production for KM1020. Bioreactor fed-batch cultivations comparing a Δspo0A abrB* mutant of KM1016, KM681, with JABs32 showed a decrease of 32% in surfactin concentration. Conclusions: The genetic differences of B. subtilis KM1016 and JABs32 give rise to new and improved fermentation methods through high cell density processes. Deletion of the spo0A locus was shown to be the reason for higher biomass concentrations. Only in combination with an elongation of abrB was this strain able to reach high surfactin titers of 18.27 g L−1 in fed-batch cultivations. This work shows, that a B. subtilis strain can be turned into a high cell density surfactin production strain by introduction of two mutations. [ABSTRACT FROM AUTHOR]

Salicornia Sinus-Persica Akhani has been discovered in southern Iran, namely in Khuzestan, Bushehr, and Hormozgan, along salty and brackish rivers draining into the Persian Gulf well as estuaries and coastal habitats. Genetic diversity is an essential component of biological variability for conservation methods, particularly for rare and narrowly endemic species. Iran's folk's genetic structure, diversity, and morphological distinctions are unknown. Because of the plant's medicinal value, genetic diversity and folk structure were studied in six regional folks of Salicornia Sinus-Persica Akhani. To emphasize genetic variability within and across people in this plant, we employed six inter-retrotransposon amplified polymorphism (IRAP) markers and 15 combined IRAP markers. The AMOVA test indicated a substantial genetic difference across the folks investigated (PhiPT = 0.41, P = 0.010), as well as the fact that within-folk variation accounted for 80% of overall genetic diversity and genetic divergence between folks accounted for 20%. The Mantel test revealed a substantial positive connection between genetic distance and geographical distance across all groups investigated. According to STRUCTURE analysis and folk assignment tests, these folks exhibit some degree of gene flow--the UPGMA classification of molecular data matched with the PCoA plot of people. Based on (IRAP) markers, these findings revealed that the regional folks of Salicornia Sinus-Persica are incredibly distinct. [ABSTRACT FROM AUTHOR]

Bacillus subtilis 3NA is a strain capable of reaching high cell densities. A surfactin producing sfp+ variant of this strain, named JABs32, was utilized in fed-batch cultivation processes. Both a glucose and an ammonia solution were fed to set a steady growth rate μ of 0.1 h-1. In this process, a cell dry weight of up to 88 g L-1 was reached after 38 h of cultivation, and surfactin titers of up to 26.5 g L-1 were detected in this high cell density fermentation process, achieving a YP/X value of 0.23 g g-1 as well as a qP/X of 0.007 g g-1 h-1. In sum, a 21-fold increase in surfactin titer was obtained compared with cultivations in shake flasks. In contrast to fed-batch operations using Bacillus subtilis JABs24, an sfp+ variant derived from B. subtilis 168, JABs32, reached an up to fourfold increase in surfactin titers using the same fed-batch protocol. Additionally, a two-stage feed process was established utilizing strain JABs32. Using an optimized mineral salt medium in this high cell density fermentation approach, after 31 h of cultivation, surfactin titers of 23.7 g L-1 were reached with a biomass concentration of 41.3 g L-1, thus achieving an enhanced YP/X value of 0.57 g g-1 as well as a qP/X of 0.018 g g-1 h-1. The mutation of spo0A locus and an elongation of AbrB in the strain utilized in combination with a high cell density fed-batch process represents a promising new route for future enhancements on surfactin production. Key points: • Utilization of a sporulation deficient strain for fed-batch operations • High cell density process with Bacillus subtilis for lipopeptide production was established • High titer surfactin production capabilities confirm highly promising future platform strain [ABSTRACT FROM AUTHOR]

Genetic variability and populations, structure were studied in seven geographical populations of Hesperis L. Genetic diversity parameters were determined in these populations. 5 of 10 random amplified polymorphic DNA primers produced 62 reproducible bands with average of 7.1 bands per primer and 55% of polymorphism. Hesperis hyrcana showed the highest number of effective allele (Ne), Shannon index (I) and genetic diversity (H). The highest values of genetic diversity were obtained in Hesperis hyrcana. NJ trees grouped the populations in two different clusters/groups, indicating their genetic difference which is discussed in details. The results of this study showed that the level of genetic variation in Hesperis is relatively high. NJ-based dendrogram showed a close relationship between members of Hesperis straussii and Hesperis hyrcana while the Hesperis luristanica protected population differ the most from the other populations. Principal component analysis, however, showed some minor differences with NJ-based dendrograms. [ABSTRACT FROM AUTHOR]

A novel approach targeting self-inducible surfactin synthesis under oxygen-limited conditions is presented. Because both the nitrate (NarGHI) and nitrite (NasDE) reductase are highly expressed during anaerobic growth of B. subtilis, the native promoter PsrfA of the surfactin operon in strain B. subtilis JABs24 was replaced by promoters PnarG and PnasD to induce surfactin synthesis anaerobically. Shake flask cultivations with varying oxygen availabilities indicated no significant differences in native PsrfA expression. As hypothesized, activity of PnarG and PnasD increased with lower oxygen levels and surfactin was not produced by PsrfA::PnarG as well as PsrfA::PnasD mutant strains under conditions with highest oxygen availability. PnarG showed expressions similar to PsrfA at lowest oxygen availability, while maximum value of PnasD was more than 5.5-fold higher. Although the promoter exchange PsrfA::PnarG resulted in a decreased surfactin titer at lowest oxygen availability, the strain carrying PsrfA::PnasD reached a 1.4-fold increased surfactin concentration with 696 mg/L and revealed an exceptional high overall YP/X of 1.007 g/g. This value also surpassed the YP/X of the reference strain JABs24 at highest and moderate oxygen availability. Bioreactor cultivations illustrated that significant cell lysis occurred when the process of "anaerobization" was performed too fast. However, processes with a constantly low agitation and aeration rate showed promising potential for process improvement, especially by employing the strain carrying PsrfA::PnasD promoter exchange. Additionally, replacement of other native promoters by nitrite reductase promoter PnasD represents a promising tool for anaerobic-inducible bioprocesses in Bacillus. [ABSTRACT FROM AUTHOR]

Strain engineering is often a method of choice towards increasing the yields of the biosurfactant surfactin which is naturally synthesized by many Bacillus spp., most notably Bacillus subtilis. In the current study, a genome reduced B. subtilis 168 strain lacking 10% of the genome was established and tested for its suitability to synthesize surfactin under aerobic and anaerobic conditions at 25 °C, 30 °C, 37 °C and 40 °C. This genome reduced strain was named IIG-Bs20-5-1 and lacks, amongst others, genes synthesizing the lipopeptide plipastatin, the antibiotic bacilysin, toxins and prophages, as well as genes involved in sporulation. Amongst all temperatures tested, 37 °C was overall superior. In comparison to the reference strain JABs24, a surfactin synthesizing variant of B. subtilis 168, strain IIG-Bs20-5-1 was both aerobically and anaerobically superior with respect to specific growth rates µ and yields YX/S. However, in terms of surfactin production, strain JABs24 reached higher absolute concentrations with up to 1147.03 mg/L and 296.37 mg/L under aerobic and anaerobic conditions, respectively. Concomitant, strain JABs24 reached higher YP/S and YP/X. Here, an outstanding YP/X of 1.541 g/g was obtained under anaerobic conditions at 37 °C. The current study indicates that the employed genome reduced strain IIG-Bs20-5-1 has several advantages over the strain JABs24 such as better conversion from glucose into biomass and higher growth rates. However, regarding surfactin synthesis and yields, the strain was overall inferior at the investigated temperatures and oxygen conditions. Further studies addressing process development and strain engineering should be performed combining the current observed advantages of the genome reduced strain to increase the surfactin yields and to construct a tailor-made genome reduced strain to realize the theoretically expected advantages of such genome reduced strains. [ABSTRACT FROM AUTHOR]

Background The objective of this randomized, double-blind, placebo-controlled study was to evaluate the efficacy of sustained-release methylphenidate (MPH-SR) in treatment of methamphetamine dependence. Methods Fifty-six individuals who met DSM-IV-TR criteria for methamphetamine dependence participated in this 10-week trial. The participants were randomly allocated into two groups and received 18 to 54 mg/day sustained-released methylphenidate or placebo for 10 weeks. Craving was evaluated by a visual analogue craving scale every week. Urinary screening test for methamphetamine was carried out each week. The Beck Depression Inventory-II (BDI-II) was used to monitor participant depressive symptoms at baseline and bi-weekly during the treatment period. Results At the end of the trial, the MPH-SR group was less methamphetamine positive compared to the placebo group and the difference was significant (p = 0.03). By the end of the study, MPH-SR group showed significantly less craving scores compared to the placebo group [MD (95% CI) = -10.28(0.88-19.18), t(54) = 2.19, p = 0.03]. There was greater improvement in the depressive symptoms scores in the intervention group compared to the placebo group [MD (95% CI) =2.03(0.31-3.75), t (54) =2.37, p = 0.02]. Conclusion Sustained-released methylphenidate was safe and well tolerated among active methamphetamine users and significantly reduced methamphetamine use, craving and depressive symptoms. [ABSTRACT FROM AUTHOR]

Background: Plipastatin is a potent Bacillus antimicrobial lipopeptide with the prospect to replace conventional antifungal chemicals for controlling plant pathogens. However, the application of this lipopeptide has so far been investigated in a few cases, principally because of the yield in low concentration and unknown regulation of biosynthesis pathways. B. subtilis synthesizes plipastatin by a non-ribosomal peptide synthetase encoded by the ppsABCDE operon. In this study, B. subtilis 3NA (a non-sporulation strain) was engineered to gain more insights about plipastatin mono-production. Results: The 4-phosphopantetheinyl transferase Sfp posttranslationally converts non-ribosomal peptide synthetases from inactive apoforms into their active holoforms. In case of 3NA strain, sfp gene is inactive. Accordingly, the first step was an integration of a repaired sfp version in 3NA to construct strain BMV9. Subsequently, plipastatin production was doubled after integration of a fully expressed degQ version from B. subtilis DSM10T strain (strain BMV10), ensuring stimulation of DegU-P regulatory pathway that positively controls the ppsABSDE operon. Moreover, markerless substitution of the comparably weak native plipastatin promoter (Ppps) against the strong constitutive promoter Pveg led to approximately fivefold enhancement of plipastatin production in BMV11 compared to BMV9. Intriguingly, combination of both repaired degQ expression and promoter exchange (Ppps::Pveg) did not increase the plipastatin yield. Afterwards, deletion of surfactin (srfAA-AD) operon by the retaining the regulatory comS which is located within srfAB and is involved in natural competence development, resulted in the loss of plipastatin production in BMV9 and significantly decreased the plipastatin production of BMV11. We also observed that supplementation of ornithine as a precursor for plipastatin formation caused higher production of plipastatin in mono-producer strains, albeit with a modified pattern of plipastatin composition. Conclusions: This study provides evidence that degQ stimulates the native plipastatin production. Moreover, a full plipastatin production requires surfactin synthetase or some of its components. Furthermore, as another conclusion of this study, results point towards ornithine provision being an indispensable constituent for a plipastatin mono-producer B. subtilis strain. Therefore, targeting the ornithine metabolic flux might be a promising strategy to further investigate and enhance plipastatin production by B. subtilis plipastatin mono-producer strains. [ABSTRACT FROM AUTHOR]